P19-2007-168 Е. Ю. Кадышевская, Н. А. Колтовая УЧАСТИЕ

P19-2007-168
…. . Š ¤ÒÏ¥¢¸± Ö, . . Šμ²Éμ¢ Ö
“—‘’ˆ… ƒ…‚ SRM5/CDC28, SRM8/NET1, SRM12/HFI1
‚ Š’ˆ‚–ˆˆ —…Š‰’-Š’‹Ÿ “ „††…‰
SACCHAROMYCES CEREVISIAE
¶· ¢²¥´μ ¢ ¦Ê·´ ² ®ƒ¥´¥É¨± ¯
Š ¤ÒÏ¥¢¸± Ö …. ., Šμ²Éμ¢ Ö . .
“Î ¸É¨¥ £¥´μ¢ SRM5/CDC28, SRM8/NET1, SRM12/HFI1
¢ ±É¨¢ ͨ¨ Î¥±¶μ°´É-±μ´É·μ²Ö Ê ¤·μ¦¦¥°
Saccharomyces cerevisiae
P19-2007-168
“ ¤·μ¦¦¥° Saccharomyces cerevisiae ¢Ò¤¥²¥´μ μ±μ²μ ¤¢ÊÌ ¤¥¸ÖÉ±μ¢ £¥´μ¢,
ÊÎ ¸É¢ÊÕÐ¨Ì ¢ Î¥±¶μ°´É-±μ´É·μ²¥. ‚ ¤ ´´μ° · ¡μÉ¥ ¨¸¸²¥¤μ¢ ²μ¸Ó ÊÎ ¸É¨¥ ¢
μ¸É ´μ¢±¥ ±²¥Éμδμ£μ ͨ±² ¶μ¤ ¤¥°¸É¢¨¥³ ¶μ¢·¥¦¤ ÕÐ¨Ì £¥´Éμ¢ £¥´μ¢ SRM,
±μÉμ·Ò¥ ¢Ò¤¥²¥´Ò · ´¥¥ ± ± £¥´Ò, ¢²¨ÖÕШ¥ ´ £¥´¥É¨Î¥¸±ÊÕ ¸É ¡¨²Ó´μ¸ÉÓ ¨
· ¤¨μÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ [1Ä3]. μ± § ´μ, ÎÉμ ³ÊÉ Í¨¨ srm5/cdc28-srm, srm8/net1srm, srm12/hˇ1-srm ¸μ±· Ð ÕÉ μ¸É ´μ¢±Ê ±²¥Éμδμ£μ ͨ±² ¶·¨ ¶μ¢·¥¦¤¥´¨¨
„Š ¨ ¢²¨ÖÕÉ ´ Î¥±¶μ°´É-μ¸É ´μ¢±Ê ¢ Ë § Ì G0/S (srm5, srm8), G1/S (srm5,
srm8, srm12), S (srm5, srm12) ¨ G2/M (srm5). μ-¢¨¤¨³μ³Ê, £¥´Ò CDC28/SRM5,
HFI1/ADA1/SRM12 ¨ NET1/SRM8 ÊÎ ¸É¢ÊÕÉ ¢ Ëμ·³¨·μ¢ ´¨¨ μɱ²¨± ±²¥É±¨ ´ ¶μ¢·¥¦¤¥´¨Ö „Š ¨ ¢²¨ÖÕÉ ´ Î¥±¶μ°´É-·¥£Ê²ÖͨÕ.
¡μÉ ¢Ò¶μ²´¥´ ¢ ‹ ¡μ· Éμ·¨¨ · ¤¨ Í¨μ´´μ° ¡¨μ²μ£¨¨ ˆŸˆ.
·¥¶·¨´É ¡Ñ¥¤¨´¥´´μ£μ ¨´¸É¨ÉÊÉ Ö¤¥·´ÒÌ ¨¸¸²¥¤μ¢ ´¨°. „Ê¡´ , 2007
Kadyshevskaya E. Yu., Koltovaya N. A.
SRM5/CDC28, SRM8/NET1 and SRM12/HFI1 Genes Participates
in Activation of Checkpoints of Yeast Saccharomyces Cerevisiae
P19-2007-168
It's known about twenty checkpoint genes in yeast Sacharomyces cerevisiae. We
are studying participation of SRM genes, selected as genes affecting genetic stability and radiosensitivity [1Ä3]. It was shown that srm5/cdc28-srm, srm8/net1-srm,
srm12/hˇ1-srm mutations prevent checkpoint activation by DNA damage, particularly
G0/S-checkpoint (srm5, srm8), G1/S-checkpoint (srm5, srm8, srm12), S-checkpoint
(srm5, srm12) and G2-checkpoint (srm5). These data indicate, at least in budding
yeast, CDC28/SRM5, HFI1/ADA1/SRM12 and NET1/SRM8 genes mediate cellular
response induced by DNA damage including checkpoint control.
The investigation has been performed at the Laboratory of Radiation Biology,
JINR.
Preprint of the Joint Institute for Nuclear Research. Dubna, 2007
‚‚…„…ˆ…
μαÊÕШ¥¸Ö ¤·μ¦¦¨ Saccharomyces cerevisiae ¸²Ê¦ É Ê¤μ¡´μ° Ôʱ ·¨μɨΥ¸±μ° ³μ¤¥²Ó´μ° ¸¨¸É¥³μ°, ¤²Ö ±μÉμ·μ° ¢¶¥·¢Ò¥ ¡Ò² · §· ¡μÉ ´ ±μ´Í¥¶Í¨Ö Î¥±¶μ°´É-·¥£Ê²Öͨ¨ ¨ ±μÉμ· Ö ¶·μ¤μ²¦ ¥É ¨¸¶μ²Ó§μ¢ ÉÓ¸Ö ¢ ³´μ£μΨ¸²¥´´ÒÌ ¨¸¸²¥¤μ¢ ´¨ÖÌ ÔÉμ£μ ³¥Ì ´¨§³ [4Ä5]. „·μ¦¦¨ ¨ ¤·Ê£¨¥ Ôʱ ·¨μɨΥ¸±¨¥ μ·£ ´¨§³Ò ¨³¥ÕÉ ´¥¸±μ²Ó±μ ÉμÎ¥± ¶·μ¢¥·±¨ (Î¥±¶μ°´ÉÒ), ¢ ±μÉμ·ÒÌ
±μ´É·μ²¨·Ê¥É¸Ö ¶· ¢¨²Ó´μ¸ÉÓ ¶·μÌ즤¥´¨Ö ±²¥Éμδμ£μ ͨ±² ¨ μ¸ÊÐ¥¸É¢²Ö¥É¸Ö μ¸É ´μ¢± ¢ μÉ¢¥É ´ ¶μ¢·¥¦¤¥´¨Ö „Š ¨²¨ ¡¥·· ´É´Ò¥ ¸É·Ê±ÉÊ·Ò
„Š, ¢μ§´¨± ÕШ¥ ¢ ¸²ÊÎ ¥ ¡²μ±¨·μ¢ ´¨Ö ·¥¶²¨± ͨ¨ „Š. —¥±¶μ°´ÉÒ,
±É¨¢¨·ÊÕШ¥¸Ö ¶·¨ ¢μ§´¨±´μ¢¥´¨¨ ¶μ¢·¥¦¤¥´¨° „Š ¢ Ë § Ì G1 ¨²¨ G2,
¶·¨¢μ¤ÖÉ ± μ¸É ´μ¢±¥ ±²¥Éμδμ£μ ͨ±² ¢ ÔÉ¨Ì ¦¥ Ë § Ì [6]. μ¢·¥¦¤¥´¨Ö „Š, ¢μ§´¨± ÕШ¥ ¢ Ë §¥ S, ¶·¨¢μ¤ÖÉ ± § ³¥¤²¥´¨Õ ·¥¶²¨± ͨ¨ „Š
¨ ± μ¸É ´μ¢±¥ ¢ Ë §¥ G2. ‡ ³¥¤²¥´¨¥ ·¥¶²¨± ͨ¨ „Š Ö¢²Ö¥É¸Ö ¸²¥¤¸É¢¨¥³
±μ³¡¨´¨·μ¢ ´´μ£μ ÔËË¥±É Î¥±¶μ°´É-´¥§ ¢¨¸¨³μ£μ ¨´£¨¡¨·μ¢ ´¨Ö ¤¢¨¦¥´¨Ö
·¥¶²¨± ɨ¢´μ° ¢¨²±¨ [7] ¨ Î¥±¶μ°´É-§ ¢¨¸¨³μ£μ ¨´£¨¡¨·μ¢ ´¨Ö μɱ·Ò¢ ´¨Ö
¸ °Éμ¢ ´ Î ² ·¥¶²¨± ͨ¨ ori [8, 9]. —¥±¶μ°´É, ¢Ò§Ò¢ ÕШ° § ³¥¤²¥´¨¥ ·¥¶²¨± ͨ¨ „Š, ´ §Ò¢ ¥É¸Ö intra S-Î¥±¶μ°´É [10, 11]. ‚Éμ·μ°, ·¥¶²¨± ɨ¢´Ò° S/M-Î¥±¶μ°´É, ËÊ´±Í¨μ´¨·ÊÕШ° ¢ Ë §¥ S, ¢Ò§Ò¢ ¥É μ¸É ´μ¢±Ê ±²¥Éμδμ£μ ͨ±² ¨ ¸Ê¶·¥¸¸¨·Ê¥É ±É¨¢ Í¨Õ ¶μ§¤´¨Ì ¸ °Éμ¢ μri-·¥¶²¨± ͨ¨ ¢ μÉ¢¥É
´ ¡²μ± ·¥¶²¨± ͨ¨ „Š, ´ ¶·¨³¥·, £¨¤·μ±¸¨³μÎ¥¢¨´μ° ¢ ·¥§Ê²ÓÉ É¥ ¨¸ÉμÐ¥´¨Ö ¤¥§μ±¸¨·¨¡μ´Ê±²¥μɨ¤É·¨Ëμ¸Ë Éμ¢ (dNTP) [9, 12]. ÉμÉ ¸¨£´ ²Ó´Ò°
¶ÊÉÓ, S-Î¥±¶μ°´É, ¶·μ¢¥·Ö¥É § ¢¥·Ï¥´¨¥ ·¥¶²¨± ͨ¨ „Š ¨ ¶·¥¤μÉ¢· Ð ¥É
³¨Éμ§ ¤μ É¥Ì ¶μ·, ¶μ± ·¥¶²¨± ꬅ ´¥ § ±μ´Î¨É¸Ö.
”Ê´±Í¨μ´ ²Ó´μ¸ÉÓ Î¥±¶μ°´Éμ¢ ±·¨É¨Î´ ¤²Ö ¶·¥¤μÉ¢· Ð¥´¨Ö £¥´μ³´μ°
´¥¸É ¡¨²Ó´μ¸É¨, ¢Ò§¢ ´´μ° ¶μ¢·¥¦¤¥´¨Ö³¨ „Š, É ± ± ± Î¥±¶μ°´ÉÒ μ¸ÊÐ¥¸É¢²ÖÕÉ § ³¥¤²¥´¨¥ ¨²¨ ·¥¸É ±²¥Éμδμ£μ ͨ±² ¢ μÉ¢¥É ´ ¶μ¢·¥¦¤¥´¨Ö „Š,
μ¡¥¸¶¥Î¨¢ Ö ¢μ§³μ¦´μ¸ÉÓ ·¥¶ · ͨ¨ [6, 11, 13, 14]. —¥±¶μ°´ÉÒ É ±¦¥ ¶·¥¤μÉ¢· Ð ÕÉ ¸¶μ´É ´´ÊÕ £¥´μ³´ÊÕ ´¥¸É ¡¨²Ó´μ¸ÉÓ [15, 16]. Ò²μ ¶μ± § ´μ, ÎÉμ
¨³¥´´μ ¤¥Ë¥±ÉÒ ·¥¶²¨± ɨ¢´μ£μ Î¥±¶μ°´É ¨ intra S-Î¥±¶μ°´É , ´μ ´¥ G1¨²¨ G2-Î¥±¶μ°´Éμ¢ ¢Ò§Ò¢ ÕÉ Ê¢¥²¨Î¥´¨¥ Ê·μ¢´Ö ¸¶μ´É ´´ÒÌ £¥´μ³´ÒÌ ¶¥·¥¸É·μ¥± [15, 16]. μ²ÊÎ¥´´Ò¥ ·¥§Ê²ÓÉ ÉÒ ¶μ§¢μ²ÖÕÉ ¶·¥¤¶μ²μ¦¨ÉÓ, ÎÉμ ±
£¥´μ³´Ò³ ¶¥·¥¸É·μ°± ³ ¶·¨¢μ¤ÖÉ μϨ¡±¨ ·¥¶²¨± ͨ¨ „Š ¨ ÎÉμ Î¥±¶μ°´ÉÒ ¢ Ë §¥ S ¸Ê¶·¥¸¸¨·ÊÕÉ Ôɨ ¶¥·¥¸É·μ°±¨.
1
•μÉÖ ·¥£Ê²Öꬅ ¶·μÌ즤¥´¨Ö ±²¥Éμδμ£μ ͨ±² ´ ¨¡μ²¥¥ Ìμ·μÏμ ¨§ÊÎ¥´ Ê ¤·μ¦¦¥° S. cerevisiae, ´μ ³´μ£¨¥ ÊÎ ¸É´¨±¨ ÔÉμ£μ ¶·μÍ¥¸¸ ¥Ð¥ ´¥
¨§¢¥¸É´Ò. ´¥¥ ´ ³¨ ¡Ò²μ ¶μ± § ´μ, ÎÉμ ³ÊÉ Í¨¨ ¢ £¥´ Ì CDC28, NET1 ¨
HFI1 ¸μ¶·μ¢μ¦¤ ÕÉ¸Ö ¶ ¤¥´¨¥³ ³¨ÉμɨΥ¸±μ° ¸É ¡¨²Ó´μ¸É¨ ¶·¨·μ¤´ÒÌ ¨ ·¥±μ³¡¨´ ´É´ÒÌ Ö¤¥·´ÒÌ £¥´¥É¨Î¥¸±¨Ì ¸É·Ê±ÉÊ· ¨ ¶μ¢ÒÏ¥´¨¥³ ÎÊ¢¸É¢¨É¥²Ó´μ¸É¨ ± ²¥É ²Ó´μ³Ê ¤¥°¸É¢¨Õ γ-¨§²ÊÎ¥´¨Ö [1, 3]. Š¨´ § CDC28, ¶μ-¢¨¤¨³μ³Ê,
¶·¨´¨³ ¥É ÊÎ ¸É¨¥ ¢ ·¥±μ³¡¨´ Í¨μ´´μ° ·¥¶ · ͨ¨, ¶μ¸±μ²Ó±Ê Ê·μ¢¥´Ó ¸¶μ´É ´´μ° £¥´´μ° ±μ´¢¥·¸¨¨ Ê ³ÊÉ ´É cdc28-srm Ê¢¥²¨Î¥´ ¢ 10 · § ¶μ ¸· ¢´¥´¨Õ
¸ ±²¥É± ³¨ ¤¨±μ£μ ɨ¶ , Î ¸ÉμÉ ¨´¤Êͨ·μ¢ ´´μ° ³¨ÉμɨΥ¸±μ° ·¥±μ³¡¨´ ͨ¨ ¢ ±²¥É± Ì ÔÉμ£μ ³ÊÉ ´É ¶μ¤ ¢²¥´ , ¶·¨ ÔÉμ³ ¸´¨¦ ¥É¸Ö ± ± Î ¸ÉμÉ £¥´´μ° ±μ´¢¥·¸¨¨, É ± ¨ ±·μ¸¸¨´£μ¢¥· [2, 17]. Š·μ³¥ Éμ£μ, ³ÊÉ Í¨Ö cdc28-srm,
ʳ¥·¥´´μ ¢²¨ÖÕÐ Ö ´ · ¤¨μÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ ±²¥Éμ±, ¶μ¢ÒÏ ¥É ¨ ¡¥§ Éμ£μ
¢Ò¸μ±ÊÕ · ¤¨μÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ ³ÊÉ ´É rad52 [2, 17]. Œμ¦´μ ¶·¥¤¶μ²μ¦¨ÉÓ,
ÎÉμ ±¨´ § ¶·¨´¨³ ¥É ÊÎ ¸É¨¥ É ±¦¥ ¢ ³¨´μ·´μ³ ¶Êɨ ·¥¶ · ͨ¨ „ „Š
¨/¨²¨ Î¥±¶μ°´É-·¥£Ê²Öͨ¨. ´ ²¨§ Ô¶¨¸É ɨΥ¸±¨Ì ¢§ ¨³μ¤¥°¸É¢¨° ³ÊÉ Í¨°
ÔÉ¨Ì £¥´μ¢ ¢ μÉ´μÏ¥´¨¨ · ¤¨μÎÊ¢¸É¢¨É¥²Ó´μ¸É¨ ¢ÒÖ¢¨² ¶·¨´ ¤²¥¦´μ¸ÉÓ £¥´μ¢
CDC28 ¨ NET1 ± μ¤´μ° RAD9-£·Ê¶¶¥ Î¥±¶μ°´É-±μ´É·μ²Ö [2, 17Ä21]. ‚¢¥¤¥´¨¥ Î¥±¶μ°´É-³ÊÉ Í¨¨ rad53 ¢ £¥´μɨ¶ ¤¢μ°´μ£μ ³ÊÉ ´É cdc28-srm rad52
´¥ ¶·¨¢μ¤¨É ± ¤μ¶μ²´¨É¥²Ó´μ³Ê Ê¢¥²¨Î¥´¨Õ γ-ÎÊ¢¸É¢¨É¥²Ó´μ¸É¨ Ê É·μ°´μ£μ
³ÊÉ ´É [21]. ‚ ´ ¸ÉμÖÐ¥° · ¡μÉ¥ ±μ¸¢¥´´Ò¥ £¥´¥É¨Î¥¸±¨¥ ¤ ´´Ò¥ μ¡ ÊÎ ¸É¨¨ £¥´ CDC28, É ±¦¥ £¥´μ¢ NET1 ¨ HFI1 ¢ Î¥±¶μ°´É-·¥£Ê²Öͨ¨ ¡Ò²¨
¶μ¤É¢¥·¦¤¥´Ò ·¥§Ê²ÓÉ É ³¨ ¶·Ö³μ£μ ´ ²¨§ § ¤¥·¦±¨ ±²¥Éμδμ£μ ͨ±² ¶μ¸²¥ ¢μ§¤¥°¸É¢¨Ö ¶μ¢·¥¦¤ ÕШ³ £¥´Éμ³. Œμ·Ëμ²μ£¨Î¥¸±¨¥ ³¥Éμ¤Ò ¶μ§¢μ²¨²¨ ¢ÒÖ¢¨ÉÓ ÊÎ ¸É¨¥ ÔÉ¨Ì £¥´μ¢ ¶· ±É¨Î¥¸±¨ ¢μ ¢¸¥Ì Î¥±¶μ°´É Ì.
1. Œ’…ˆ‹› ˆ Œ…’„›
˜É ³³Ò ³¨±·μμ·£ ´¨§³μ¢. ‚¸¥ ÏÉ ³³Ò ¤·μ¦¦¥° Saccharomyces cerevisiae, ¨¸¶μ²Ó§μ¢ ´´Ò¥ ¢ ¤ ´´μ° · ¡μÉ¥, ¢ Éμ³ Î¨¸²¥ 711a (MATa SRM +
ade2), M1a (MATa srm1 ade1), M5a (MATa srm5 ade1), C3 (MATa srm8 leu2
ade2), 9a (MATa srm12 ade2 leu2 ura3 trp1), 3C (MATa rad9::LEU2 ade2),
1D (MATα rad17::URA3 ade2), 16B (MATa rad24::URA3 ade2), 14α (MATα
srm12 ade2 leu2 ura3), 12C (MAT rad53 ade2) Å ¸±μ´¸É·Ê¨·μ¢ ´Ò ¸μ¢³¥¸É´μ
¸ ¤-·μ³ . . „¥¢¨´Ò³ (ˆŒƒ , Œμ¸±¢ ) ´ £¥´μËμ´¥ ÏÉ ³³ 71a [2].
‚¢¥¤¥´¨¥ ³ ·±¥·μ¢ ¶·μ¨§¢μ¤¨²μ¸Ó ¸ ¶μ³μÐÓÕ £¥´¥É¨Î¥¸±¨Ì ³¥Éμ¤μ¢ c ¨¸¶μ²Ó§μ¢ ´¨¥³ ³¨±·μ³ ´¨¶Ê²ÖÉμ· . ‘±μ´¸É·Ê¨·μ¢ ´´Ò¥ ÏÉ ³³Ò ´¥¸ÊÉ ³ÊÉ Í¨¨ srm1, srm5/cdc28-srm [1], srm8/net1-srm ¨ srm12/hˇ1-srm [3]. ‚ ± Î¥¸É¢¥ ¨¸Éμ䨱 ³ÊÉ Í¨° rad9Δ, rad17Δ, rad24Δ ¨ rad53 ¸²Ê¦¨²¨ ÏÉ ³³Ò:
7859-7-4a (MATa rad9::LEU2 leu2-3,112 trp1-289 ura3-52 his7), ²Õ¡¥§´μ ¶·¨¸² ´´Ò° ´ ³ Prof. L. H. Hartwell (University of Washington, Seattle); SX46A
rad24Δ (MATa rad24::URA3 ade2 his3-532 trp1-289 ura3-52), SX46 rad17Δ
2
(MATa rad17::URA3 ade2 his3-532 trp1-289 ura3-52) ¨ CRY1 (MATa sad1-1
(=rad53) ade2-1 ura3-1 trp1-1 his3-11,15 leu2-3,112 can1-100), ¶μ²ÊÎ¥´´Ò¥ μÉ
Prof. W. Siede (University of Texas, Dallas). ‚ ± Î¥¸É¢¥ ¨¸Éμ䨱 ³ÊÉ Í¨°
cdc6-1 ¨ cdc9-1 ¸²Ê¦¨²¨ ÏÉ ³³Ò 327A (MATa cdc6-1 ade1 ade2 ura1 his7 lys2
tyr1 gal1) ¨ STX435-1-3B (MATα cdc9-1 ade1 ade2 lys2 ura1 his7 leu1 gal1),
¶μ²ÊÎ¥´´Ò¥ μÉ ¤-· . ˆ. ¢²μ¢ (‘ƒ“, ‘.-¥É¥·¡Ê·£).
‘·¥¤Ò ¨ ·¥ ±É¨¢Ò. ˆ¸¶μ²Ó§μ¢ ²¨ ¸É ´¤ ·É´ÊÕ ¶μ²´ÊÕ ¶¨É É¥²Ó´ÊÕ ¸·¥¤Ê
YPD [22] ¨ ¶μ²´ÊÕ ¸·¥¤Ê ‘, 춨¸ ´´ÊÕ · ´¥¥ [1]. ‘¨´Ì·μ´¨§ Í¨Õ ±Ê²ÓÉÊ·
μ¸ÊÐ¥¸É¢²Ö²¨ ¸ ¶μ³μÐÓÕ α-Ë ±Éμ· S. cerevisiae (Peptide Institute, Inc). ‚
· ¡μÉ¥ ¨¸¶μ²Ó§μ¢ ²¨ É ±¦¥ £¨¤·μ±¸¨³μÎ¥¢¨´Ê (Sigma).
“”-¸¢¥É. „²Ö μ¡²ÊÎ¥´¨Ö ¢Ò· Ш¢ ²¨ ´μδҥ ±Ê²ÓÉÊ·Ò ¤·μ¦¦¥° ¢ ¦¨¤±μ° ¶¨É É¥²Ó´μ° ¸·¥¤¥ YPD ¶·¨ 30 ◦ ‘. Š²¥É±¨ · §¢μ¤¨²¨ ¢ ¢μ¤¥, ´ ´μ¸¨²¨
´ ¶μ¢¥·Ì´μ¸ÉÓ Î Ï±¨ ¸ £ ·¨§μ¢ ´´μ° ¸·¥¤μ° ‘ ¨§ · ¸Î¥É 100 ¦¨§´¥¸¶μ¸μ¡´ÒÌ ±²¥Éμ± ´ Î Ï±Ê ¨ μ¡²ÊÎ ²¨ ¢ É¥³´μÉ¥ “”-¸¢¥Éμ³ (² ³¶ „
30 w,
³μдμ¸ÉÓ ¤μ§Ò 0,3 „¦/³2 ¸). ¡²ÊÎ¥´´Ò¥ Πϱ¨ ¶μ³¥Ð ²¨ ¢ ¶¥´ ² ¢μ ¨§¡¥¦ ´¨¥ ËμÉμ·¥ ±É¨¢ ͨ¨ ¨ ¢³¥¸É¥ ¸ ±μ´É·μ²Ó´Ò³¨ Πϱ ³¨ ¨´±Ê¡¨·μ¢ ²¨ ¢
ɥΥ´¨¥ 5 ¸ÊÉ ¶·¨ 30 ◦ ‘.
‘¨´Ì·μ´¨§ ꬅ ±Ê²ÓÉÊ· ¢ Ë §¥ G1 ¸ ¶μ³μÐÓÕ α-Ë ±Éμ· . ƒ ¶²μ¨¤´Ò¥
±²¥É±¨ ɨ¶ ¸¶ ·¨¢ ´¨Ö α ¸¨´É¥§¨·ÊÕÉ α-Ë ±Éμ·, ±μÉμ·Ò° ¢Ò¤¥²Ö¥É¸Ö ¢ ¸·¥¤Ê
¨ μ± §Ò¢ ¥É ¢²¨Ö´¨¥ ´ ±²¥É±¨ ¶·μɨ¢μ¶μ²μ¦´μ£μ ɨ¶ ¸¶ ·¨¢ ´¨Ö . μ¤
¢μ§¤¥°¸É¢¨¥³ ÔÉμ£μ Ë ±Éμ· ±²¥É±¨ μ¸É ´ ¢²¨¢ ÕÉ¸Ö ´ ¸É ¤¨¨ ¨´¨Í¨ ͨ¨
¸¨´É¥§ „Š. α-Ë ±Éμ· ¨¸¶μ²Ó§ÊÕÉ ¤²Ö ¸¨´Ì·μ´¨§ ͨ¨ ±Ê²ÓÉÊ· ¢ Ë §¥ G1.
„²Ö ÔÉμ£μ £ ¶²μ¨¤´Ò¥ ±Ê²ÓÉÊ·Ò ¸ £¥´μɨ¶μ³ MATa ¢Ò· Ш¢ ²¨ ¢ ɥΥ´¨¥
´μΨ ¢ ¦¨¤±μ° ¶¨É É¥²Ó´μ° ¸·¥¤¥ YPD ¤μ ±μ´Í¥´É· ͨ¨ 2−5·106 ±²/³² (· ´´ÖÖ Ô±¸¶μ´¥´Í¨ ²Ó´ Ö ±Ê²ÓÉÊ· ) ¶·¨ 30 ◦ ‘. Š²¥É±¨ μ¸ ¦¤ ²¨ Í¥´É·¨ËÊ£¨·μ¢ ´¨¥³ (1500 g, 3 ³¨´) ¨ ·¥¸Ê¸¶¥´¤¨·μ¢ ²¨ ¤μ ±μ´Í¥´É· ͨ¨ 3·107 ±²./³²
¢ 1 ³² ¸¢¥¦¥° ¶¨É É¥²Ó´μ° ¸·¥¤Ò YPD (pH 4,0), ¸μ¤¥·¦ Ð¥° 5 ³±£/³² αË ±Éμ· S. cerevisiae. μ¸²¥ ¨´±Ê¡ ͨ¨ 1,5 Î ¶·¨ 30 ◦ ‘ ¤μ¡ ¢²Ö²¨ ¢Éμ·ÊÕ
¶μ·Í¨Õ α-Ë ±Éμ· ¨ ¨´±Ê¡ Í¨Õ ¶·μ¤μ²¦ ²¨ ¥Ð¥ 1,5 Î. ‘É¥¶¥´Ó ¸¨´Ì·μ´¨§ ͨ¨ ±μ´É·μ²¨·μ¢ ²¨ ³¨±·μ¸±μ¶¨·μ¢ ´¨¥³ ±Ê²ÓÉÊ·Ò, μÍ¥´¨¢ Ö ±μ²¨Î¥¸É¢μ
¶μαÊÕÐ¨Ì¸Ö ±²¥Éμ±. ‘¨´Ì·μ´¨§μ¢ ´´Ò¥ ±²¥É±¨ ¨³¥²¨ Ì · ±É¥·´ÊÕ μ¢ ²Ó´ÊÕ Ëμ·³Ê.
“”-μ¡²ÊÎ¥´¨¥ ±Ê²ÓÉÊ·, ¸¨´Ì·μ´¨§μ¢ ´´ÒÌ ¢ Ë §¥ G1, ¸ ¶μ³μÐÓÕ
α-Ë ±Éμ· . ‘¨´Ì·μ´¨§μ¢ ´´ÊÕ α-Ë ±Éμ·μ³ ±Ê²ÓÉÊ·Ê μ¸ ¦¤ ²¨ Í¥´É·¨ËÊ£¨·μ¢ ´¨¥³ (1500 g, 3 ³¨´), μɳҢ ²¨ ¢ μ̲ ¦¤¥´´μ° ¢μ¤¥ ¨ ·¥¸Ê¸¶¥´§¨·μ¢ ²¨
¢ μ̲ ¦¤¥´´μ° ¢μ¤¥ ¤μ ±μ´Í¥´É· ͨ¨ 2, 5·107 ±²./³². ŠÊ²ÓÉÊ·Ê · ¸¸¥¢ ²¨ ´ Πϱ¨ ¥É·¨ ¨ ´¥³¥¤²¥´´μ μ¡²ÊÎ ²¨ “”-¸¢¥Éμ³ (¸³. “”-μ¡²ÊÎ¥´¨¥). ɳ¥É¨³, ÎÉμ ¤²Ö ¤μ¸É¨¦¥´¨Ö ´ ²μ£¨Î´μ£μ ¡¨μ²μ£¨Î¥¸±μ£μ ÔËË¥±É ¶·¨ μ¡²ÊÎ¥´¨¨ ±²¥Éμ± ¢ ¸Ê¸¶¥´§¨¨ É·¥¡ÊÕÉ¸Ö ¤μ§Ò ¶·¨³¥·´μ ¢ É·¨ · § ¡μ²ÓϨ¥, Î¥³
¶·¨ μ¡²ÊÎ¥´¨¨ ´ ¶μ¢¥·Ì´μ¸É¨ Πϥ±. ˆ¸¶μ²Ó§Ê¥³Ò¥ ´ ³¨ ±Ê²ÓÉÊ·Ò ´¥ μÉ´μ¸ÖÉ¸Ö ± ¸¨²Ó´μ ±μ³±ÊÕШ³¸Ö. „²Ö ¸´¨¦¥´¨Ö ±μ³±μ¢ ´¨Ö ¨ · ¸Ì즤¥´¨Ö
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¢ ¸¨´Ì·μ´¨§μ¢ ´´ÒÌ ±Ê²ÓÉÊ· Ì ±²¥Éμ± · §´μ£μ £¥´μɨ¶ Œ¥Éμ¤
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¢ ʸ²μ¢¨ÖÌ ¨´É¥´¸¨¢´μ° Ô· ͨ¨ ¶·¨ 30 ◦ ‘
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¨ srm12 ¸¨²Ó´μ Ê¢¥²¨Î¨¢ ²¨ ¢·¥³Ö £¥´¥· ͨ¨, ³ÊÉ ´ÉÒ ´ Ψ´ ²¨ ¤¥²¨ÉÓ¸Ö
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¢ Ë §¥ G1 Ê ³ÊÉ ´Éμ¢ srm5, srm8 ¨ srm12 ¨ ¢ Ë §¥ G0 Ê ³ÊÉ ´Éμ¢ srm5 ¨
srm8. ŒÊÉ Í¨¨ srm5 ¨ srm8, ¶μ-¢¨¤¨³μ³Ê, ¢²¨ÖÕÉ É ±¦¥ ´ intra S-Î¥±¶μ°´É,
§ ³¥¤²ÖÕШ° ¸¨´É¥§ „Š.
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¤¥°¸É¢¨¥³ £¨¤·μ±¸¨³μÎ¥¢¨´Ò
ƒ¨¤·μ±¸¨³μÎ¥¢¨´ (HU) ¨´£¨¡¨·Ê¥É Ë¥·³¥´É ·¨¡μ´Ê±²¥μɨ¤·¥¤Ê±É §Ê ¨
¶·¨¢μ¤¨É ± ´¥¸¶μ¸μ¡´μ¸É¨ ¸¨´É¥§¨·μ¢ ÉÓ ¤¥§μ±¸¨·¨¡μ´Ê±²¥μɨ¤Ò ¨ ¡²μ±Ê
¸¨´É¥§ „Š. Š²¥É±¨ ¤¨±μ£μ ɨ¶ ¢ ¶·¨¸Êɸɢ¨¨ HU μ¡· ɨ³μ μ¸É ´ ¢²¨¢ ÕÉ ¤¥²¥´¨¥ ¢ Ë §¥ S, ±É¨¢¨·ÊÖ ·¥¶²¨± ɨ¢´Ò° S-Î¥±¶μ°´É [27]. ŒÊÉ Í¨¨
8
¶μ S-Î¥±¶μ°´ÉÊ ¤μ²¦´Ò ¶·μÖ¢²ÖÉÓ ÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ ± HU, ¶μ¸±μ²Ó±Ê ´¥¸¶μ¸μ¡´μ¸ÉÓ § ¤¥·¦¨¢ ÉÓ ¤¥²¥´¨¥ ³μ¦¥É ¶·¨¢¥¸É¨ ± ³¨Éμ§Ê ¢ ʸ²μ¢¨ÖÌ ´¥§ ¢¥·Ï¥´´μ£μ ¸¨´É¥§ „Š.
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0,2 Œ HU ¨ ¨´±Ê¡¨·ÊÖ ¢ ɥΥ´¨¥ 2 ¸ÊÉ. 쳨³μ ³ÊÉ Í¨° srm ¤²Ö ¸· ¢´¥´¨Ö
´ ²¨§¨·μ¢ ²¨ É ±¦¥ ³ÊÉ Í¨¨ ¢ ¨§¢¥¸É´ÒÌ Î¥±¶μ°´É-£¥´ Ì rad9Δ, rad17Δ,
rad24Δ ¨ rad53. ¥§Ê²ÓÉ ÉÒ É¥¸É¨·μ¢ ´¨Ö £ ¶²μ¨¤´ÒÌ ÏÉ ³³μ¢ 711a (SRM + ),
M1a (srm1), M5a (srm5), C3 (srm8), 9a (srm12), 12C (rad53), 16B (rad24Δ), 1D
(rad17Δ), 3C (rad9Δ) ¶·¨¢¥¤¥´Ò ´ ·¨¸. 2, . ‚ ¶·¨¸Êɸɢ¨¨ 0,2 Œ HU ±²¥É±¨
¤¨±μ£μ ɨ¶ · ¸ÉÊÉ, ÎÉμ ¸μ£² ¸Ê¥É¸Ö ¸ ¤ ´´Ò³¨ ¤·Ê£¨Ì ¢Éμ·μ¢ [28]. ŒÊÉ Í¨¨
rad53, srm8 ¨ srm12 ¶μ¢ÒÏ ÕÉ ÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ ± HU ¢ ʸ²μ¢¨ÖÌ ·μ¸É ´ ¸·¥¤¥ ¸ 0,2 Œ HU. ŒÊÉ Í¨¨ ³μ¦´μ ¢Ò¸É·μ¨ÉÓ ¢ ·Ö¤ ¶μ ³¥·¥ ¢μ§· ¸É ´¨Ö
ÎÊ¢¸É¢¨É¥²Ó´μ¸É¨ ± HU:
SRM + <srm5<rad9<srm1<rad24<srm8<rad17<srm12<rad53.
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srm, ¤·Ê£¨³¨ Ë ±Éμ· ³¨, ²μ± ²¨§μ¢ ´´Ò³¨ ¢ ÔÉ¨Ì ÏÉ ³³ Ì. „²Ö ¶·μ¢¥·±¨
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¶μ¸É³¥°μɨΥ¸±¨Ì ¶μÉμ³±μ¢ £¨¡·¨¤μ¢ SRM+ × srm8 ¨ SRM+ × srm12. ’ ± ± ±
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(·¨¸. 2, ¡ ). ‘¥£·¥£ ´ÉÒ srm12 μɲ¨Î ÕÉ¸Ö μÉ ±²¥Éμ± ¤¨±μ£μ ɨ¶ ³¥¤²¥´´Ò³
·μ¸Éμ³. ‘· ¢´¥´¨¥ μɶ¥Î É±μ¢ ±²μ´μ¢ ´ ¸·¥¤ Ì ‘ ¨ ‘ + 0, 2 Œ HU ¤¥³μ´¸É·¨·Ê¥É · ¸Ð¥¶²¥´¨¥ ¶μ ÎÊ¢¸É¢¨É¥²Ó´μ¸É¨ ± HU ¢ μÉ´μÏ¥´¨¨ 2 : 2, ¶·¨Î¥³
ÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ ± HU ±μ··¥²¨·Ê¥É ¸ ¶·¨¸Êɸɢ¨¥³ ¢ ±²¥É± Ì ³ÊÉ Í¨° srm8
¨²¨ srm12.
„²Ö ±μ²¨Î¥¸É¢¥´´μ° μÍ¥´±¨ ÎÊ¢¸É¢¨É¥²Ó´μ¸É¨ ± ²¥É ²Ó´μ³Ê ¤¥°¸É¢¨Õ
HU ¨¸¶μ²Ó§μ¢ ²¨ £ ¶²μ¨¤´Ò¥ ÏÉ ³³Ò -ɨ¶ ¸¶ ·¨¢ ´¨Ö 711 (SRM + ), Œ5 (srm5), C3 (srm8), 9a (srm12), 3C (rad9Δ), 12C (rad53). ¡· ¡μɱ · ¸ÉÊÐ¥°
±Ê²ÓÉÊ·Ò HU ¶·¨¢μ¤¨É ± ¨Ì ¸¨´Ì·μ´¨§ ͨ¨ ¢ Ë §¥ S (0,2 Œ HU, 150 ³¨´),
Ê Î¥±¶μ°´É-³ÊÉ ´Éμ¢ É ±μ° ¸¨´Ì·μ´¨§ ͨ¨ ´¥ ¶·μ¨¸Ì줨É. ´ ²¨§ ±¨´¥É¨±¨
²¥É ²Ó´μ£μ ¤¥°¸É¢¨Ö 0,2 Œ HU ´ ¸¨´Ì·μ´´Ò¥ ±Ê²ÓÉÊ·Ò ³ÊÉ ´É´ÒÌ ±²¥Éμ±
¶μ± § ², ÎÉμ Éμ²Ó±μ ³ÊÉ Í¨Ö rad53 ¢²¨Ö¥É ´ ÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ ± ²¥É ²Ó´μ³Ê
¤¥°¸É¢¨Õ 0,2 Œ HU (·¨¸. 2, ¢). “ ÏÉ ³³μ¢ ¤¨±μ£μ ɨ¶ μ¸É ´μ¢± ¤¥²¥´¨Ö
¶·μ¤μ²¦ ¥É¸Ö ¢ ɥΥ´¨¥ 6 Î, § É¥³ ±²¥É±¨ ´ Ψ´ ÕÉ ¤¥²¨ÉÓ¸Ö. ‚ ÔÉ¨Ì Ê¸²μ¢¨ÖÌ ³ÊÉ Í¨¨ srm5 ¨ srm12 ´¥ ¢²¨Ö²¨ ´ ÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ ± HU. ¤´ ±μ
¸¨´Ì·μ´¨§ ꬅ ±Ê²ÓÉÊ·Ò ±²¥Éμ± α-Ë ±Éμ·μ³ ¢ Ë §¥ G1 ¨ ¶μ¢ÒÏ¥´¨¥ ±μ´Í¥´É· ͨ¨ ¤μ 0,5 Œ HU ¢ÒÖ¢¨²μ ¢²¨Ö´¨¥ ³ÊÉ Í¨¨ srm12 ´ ÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ
± ²¥É ²Ó´μ³Ê ¤¥°¸É¢¨Õ HU (·¨¸. 2, ¢).
¤´ ±μ μ¸É ´μ¢± ¤¥²¥´¨Ö ±²¥Éμ± ¢ ¶·¨¸Êɸɢ¨¨ HU ´¥ Ô±¢¨¢ ²¥´É´ HU-¨´¤Êͨ·μ¢ ´μ° £¨¡¥²¨ [27]. ˆ§ ·¨¸. 4 ¢¨¤´μ, ÎÉμ ±²¥É±¨ ¤¨±μ£μ ɨ¶ 9
¨¸. 2. —Ê¢¸É¢¨É¥²Ó´μ¸ÉÓ ±²¥Éμ± ± £¨¤·μ±¸¨³μÎ¥¢¨´¥. ) ɶ¥Î ɱ¨ ±²¥Éμ± · §´μ£μ
£¥´μɨ¶ (1-° ·Ö¤ Å SRM + , 2-° ·Ö¤ Å srm1, 3-° ·Ö¤ Å srm5, 4-° ·Ö¤ Å srm8, 5-°
·Ö¤ Å srm12, 6-° ·Ö¤ Å rad53, 7-° ·Ö¤ Å rad24, 8-° ·Ö¤ Å rad17, 9-° ·Ö¤ Å rad9)
´ Πϱ Ì ¸μ ¸·¥¤ ³¨ ‘+0,2 M HU (¸²¥¢ ) ¨ ‘ (¸¶· ¢ ). ¡) ’¥É· ¤Ò, ¶μ²ÊÎ¥´´Ò¥
μÉ £¨¡·¨¤μ¢ SRM+ × srm8 (¢¢¥·ÌÊ) ¨ SRM+ × srm12 (¢´¨§Ê), ¶¥·¥´¥¸¥´Ò ·¥¶²¨± Éμ·μ³ ´ ¸·¥¤Ò ‘ (30 ¨ 37 ◦ ‘) ¨ ‘+0,2 M HU. ¢) —Ê¢¸É¢¨É¥²Ó´μ¸ÉÓ ±²¥Éμ± · §´μ£μ
£¥´μɨ¶ ± ²¥É ²Ó´μ³Ê ¤¥°¸É¢¨Õ HU. Š²¥É±¨ ¢Ò· Ш¢ ²¨ ¢ ¶¨É É¥²Ó´μ° ¸·¥¤¥, ¶¥·¥´μ¸¨²¨ ¢ ¸¢¥¦ÊÕ ¶¨É É¥²Ó´ÊÕ ¸·¥¤Ê ¸ 0,2 Œ HU, Î¥·¥§ μ¶·¥¤¥²¥´´Ò¥ ¶·μ³¥¦Êɱ¨
¢·¥³¥´¨ μɳҢ ²¨ μÉ HU ¨ · ¸¸¥¢ ²¨ ´ ¶¨É É¥²Ó´ÊÕ ¸·¥¤Ê (¸²¥¢ ). ¥·¥¤ μ¡· ¡μɱμ°
0,5 Œ HU ±²¥É±¨ ¸¨´Ì·μ´¨§μ¢ ²¨ α-Ë ±Éμ·μ³ ¢ Ë §¥ G1 (¸¶· ¢ )
10
¨ ³ÊÉ ´É srm5 ¸¶μ¸μ¡´Ò · ¸É¨ ¢ ¶·¨¸Êɸɢ¨¨ 0,2 Œ HU. ˆ´£¨¡¨·μ¢ ´¨¥
¤¥²¥´¨Ö ´ ¡²Õ¤ ¥É¸Ö ²¨ÏÓ ¢ ɥΥ´¨¥ ´¥¸±μ²Ó±¨Ì Î ¸μ¢ (·¨¸. 2, ¢), ¶μÉμ³ ¶·μ¨¸Ìμ¤¨É ¤ ¶É ͨÖ, ¨ ±²¥É±¨ ´ Ψ´ ÕÉ ¤¥²¨ÉÓ¸Ö. ˆ¸¶μ²Ó§μ¢ ´¨¥ ³¥Éμ¤ ³¨±·μ±μ²μ´¨° ¶μ§¢μ²Ö¥É μ¶·¥¤¥²¨ÉÓ ¢²¨Ö´¨¥ ³ÊÉ Í¨° srm ´ § ¤¥·¦±Ê ¤¥²¥´¨Ö
¶μ¤ ¤¥°¸É¢¨¥³ HU. ·¥¸É ±²¥Éμ± μÍ¥´¨¢ ²¨ ¶·¨ μ¡· ¡μɱ¥ ±²¥Éμ± 0,2 Œ HU
¢ ɥΥ´¨¥ 4 Î. ‚ É ¡². 2 ¶·¨¢¥¤¥´Ò ·¥§Ê²ÓÉ ÉÒ É·¥Ì Ô±¸¶¥·¨³¥´Éμ¢.
’ ¡²¨Í 2. ·¥¸É ¨ £¨¡¥²Ó ±²¥Éμ± · §´μ£μ £¥´μɨ¶ ¶μ¤ ¤¥°¸É¢¨¥³ 0,2 Œ £¨¤·μ±¸¨³μÎ¥¢¨´Ò (4 Î)
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III
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67,8
12,2
37,3
83,7
62,7
16,3
37,4±5,3
Å
85,4±2,1 93,0±4,4
SRM+
−
+
29,1
39,7
70,9
60,3
24,6
37,4
75,4
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22,0
34,2
78,0
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37,1±2,8
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−
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27,0
30,6
73,0
69,4
32,3
36,1
67,7
63,9
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27,6
70,2
72,4
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Å
31,4±4,3 35,1±9,6
srm12
−
+
31,4
41,8
68,6
58,2
32,1
22,2
67,9
77,8
31,3
30,1
68,7
69,9
31,6±0,4
Å
31,4±9,9 37,1±18,3
1Ä6
Å
3,4±3,4
´μ³¥· Ô±¸¶¥·¨³¥´É ; ¡ ±μ²¨Î¥¸É¢μ ±²¥Éμ± ¢ ³¨±·μ±μ²μ´¨¨; ¢ ±μ²¨Î¥¸É¢μ (%) ³¨±·μ±μ²μ´¨° ¤ ´´μ£μ · §³¥· ; £ ¤μ²Ö ³¨±·μ±μ²μ´¨° · §³¥·μ³ 1Ä6 ±²¥Éμ±; ¤ μ¶·¥¤¥²Ö²¨ ¨§
±·¨¢ÒÌ ¢Ò¦¨¢ ¥³μ¸É¨ ¶·¨ μ¡· ¡μɱ¥ 0,2 Œ HU (4 Î)
‚¨¤´μ, ÎÉμ Ê ³ÊÉ ´É rad53 0,2 Œ HU ¢Ò§Ò¢ ¥É ´ ±μ¶²¥´¨¥ ±μ²μ´¨°
· §³¥·μ³ 1Ä6 ±²¥Éμ±. ‘μ¶μ¸É ¢²¥´¨¥ ¸ ²¥É ²Ó´μ¸ÉÓÕ ¶μ± §Ò¢ ¥É, ÎÉμ ÔÉμ ¶μ£¨¡Ï¨¥ ±²¥É±¨, É. ¥. ¶μ²´μ¸ÉÓÕ μɸÊÉ¸É¢Ê¥É § ¤¥·¦± . ’ ±¨³ μ¡· §μ³, ¢Ò¤¥·¦¨¢ ´¨¥ ±²¥Éμ± rad53 ¢ ¶·¨¸Êɸɢ¨¨ HU ¢Ò§Ò¢ ¥É ¡Ò¸É·ÊÕ £¨¡¥²Ó ±²¥Éμ±.
μ± § ´μ, ÎÉμ ¶·¨ ÔÉμ³ ´ ± ¶²¨¢ ÕÉ¸Ö ¶μαÊÕШ¥¸Ö ±²¥É±¨, ¸μ¤¥·¦ Ш¥
³¨ÉμɨΥ¸±μ¥ ¢¥·¥É¥´μ ¨ G1-„Š [27]. “ ±²¥Éμ± ¤¨±μ£μ ɨ¶ ±μ²¨Î¥¸É¢μ
³¨±·μ±μ²μ´¨° (1Ä6 ±²¥Éμ±) ¢ÒÏ¥ ¢ ¶·¨¸Êɸɢ¨¨ HU. Éμ ¶·¥¢ÒÏ¥´¨¥ ³μ¦¥É ¡ÒÉÓ μ¡Ê¸²μ¢²¥´μ ¸Ê³³ ·´Ò³ ÔËË¥±Éμ³ μ¡· ɨ³μ° ¨ ´¥μ¡· ɨ³μ° μ¸É ´μ¢±¨ ±²¥Éμδμ£μ ͨ±² . μ± § ´μ, ÎÉμ ¡μ²ÓÏ Ö Î ¸ÉÓ ¸¨´É¥§ „Š ³μ¦¥É
ÔËË¥±É¨¢´μ ¢μ¸¸É ´ ¢²¨¢ ÉÓ¸Ö ¶μ¸²¥ HU-¨´£¨¡¨·μ¢ ´¨Ö [27]. “ ³ÊÉ ´Éμ¢
srm5 ¨ srm12 ±²¥É±¨ ¤¥²ÖÉ¸Ö μ¤¨´ ±μ¢μ ¢´¥ § ¢¨¸¨³μ¸É¨ μÉ ¶·¨¸Êɸɢ¨Ö HU.
“ ÏÉ ³³ ¤¨±μ£μ ɨ¶ ±²¥É±¨ ¢ ³¨±·μ±μ²μ´¨ÖÌ ¢ ¡μ²ÓϨ´¸É¢¥ ¸¢μ¥³ ¸¶μ¸μ¡´Ò μ¡· ɨ³μ ¤¥²¨ÉÓ¸Ö, Ê ³ÊÉ ´Éμ¢ ¦¥ srm5 ¨ srm12 ·μ¸É ³¨±·μ±μ²μ´¨°
¡²μ±¨·μ¢ ´ ´¥μ¡· ɨ³μ. ‹¥É ²Ó´Ò° ÔËË¥±É ³ÊÉ Í¨° srm5 ¨ srm12 ³¥´ÓÏ¥,
Î¥³ Ê ³ÊÉ ´É rad53, ¢¥²¨Î¨´ ±μÉμ·μ£μ Ìμ·μÏμ ¸μ£² ¸Ê¥É¸Ö ¸ · ´¥¥ ¶μ²Ê11
Î¥´´Ò³¨ ¤ ´´Ò³¨ (·¨¸. 2, ¢). “ ³ÊÉ ´Éμ¢ μɸÊÉ¸É¢Ê¥É μ¡· ɨ³ Ö ±μ³¶μ´¥´É ,
¶μ-¢¨¤¨³μ³Ê, ³ÊÉ Í¨¨ ´ ·ÊÏ ÕÉ μ¸É ´μ¢±Ê ¤¥²¥´¨Ö ¢ ¶·¨¸Êɸɢ¨¨ HU ¨ É¥³
¸ ³Ò³ ¶·¨¢μ¤ÖÉ ± ¶μ¢ÒÏ¥´¨Õ ²¥É ²Ó´μ£μ ÔËË¥±É .
μ¸±μ²Ó±Ê ɨ³¨¤¨´μ¢Ò¥ ¤¨³¥·Ò, ¨´¤Êͨ·μ¢ ´´Ò¥ ¶μ¤ ¤¥°¸É¢¨¥³ “”¸¢¥É , É ±¦¥ ¢Ò§Ò¢ ÕÉ ¡²μ± ·¥¶²¨± ͨ¨ [28], ³Ò ¨§ÊÎ ²¨ ¸¶μ¸μ¡´μ¸ÉÓ ³ÊÉ Í¨° srm μ¶μ¸·¥¤μ¢ ÉÓ ¢Ò¦¨¢ ¥³μ¸ÉÓ ¢ μÉ¢¥É ´ μ¡²ÊÎ¥´¨¥ “”-¸¢¥Éμ³. ˆ§
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ͨ±² ¶·¨ ¶μ¢·¥¦¤¥´¨¨ „Š ¶μ¤ ¤¥°¸É¢¨¥³ γ-μ¡²ÊÎ¥´¨Ö.
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Î¥±¶μ°´ÉÊ Ì · ±É¥·´ £¥´¥É¨Î¥¸± Ö ´¥¸É ¡¨²Ó´μ¸ÉÓ [15, 16]. ´¥¥ ´ ³¨ ¡Ò²μ
¶μ± § ´μ, ÎÉμ £¥´Ò CDC28, NET1, HFI1 ¶·¨´¨³ ÕÉ ÊÎ ¸É¨¥ ¢ ¸É ¡¨²¨§ ͨ¨ · §²¨Î´ÒÌ £¥´¥É¨Î¥¸±¨Ì ¸É·Ê±ÉÊ·, É ±¨Ì ± ± ¶·¨·μ¤´Ò¥ Ì·μ³μ¸μ³Ò ¨
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¨¸. 6. Š¨´¥É¨± ·μ¸É ³¨±·μ±μ²μ´¨° Ê £ ¶²μ¨¤´ÒÌ ±²¥Éμ±, μ¡²ÊÎ¥´´ÒÌ ¢ Ô±¸¶μ´¥´Í¨ ²Ó´μ° Ë §¥ ·μ¸É ¢ ¤μ§ Ì 0, 20 ¨ 40 ƒ·: SRM + ( ), rad9 (¡), srm5 (¢), srm8 (£),
srm12 (¤)
18
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40
29, 37, 34
60, 51, 45
27, 36, 37
62, 60, 60
1,01±0,04
0,86±0,06
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40
37, 27, 25
49, 53, 49, 44
36, 26, 32
69, 61, 53, 42
0,94±0,06
0,89±0,07
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20
40
46, 32, 32
64, 66, 62
56, 45, 46
82, 70, 81
0,74±0,04
0,83±0,06
srm12
20
40
33, 29, 34
70, 59, 55
40, 30, 32
62, 62, 61
0,95±0,07
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20
40
8, 10, 10
18, 12
61, 59, 59
87, 80
0,16±0,0
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rad17Δ, rad24Δ ¨ rad53 ¨ μÉμ¡· ²¨ ¶μ ¤¢¥ ɥɷ ¤Ò ¶μ¸É³¥°μɨΥ¸±¨Ì ¶μÉμ³±μ¢. ‚ ±Ê²ÓÉÊ· Ì ¸¥£·¥£ ´Éμ¢ μ¶·¥¤¥²Ö²¨ Î ¸ÉμÉÊ ³ÊÉ ´Éμ¢ rho− . Š ±
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SRM5/CDC28, SRM8/NET1, SRM12/HFI1 ¢ · ¡μÉ¥ Î¥±¶μ°´É-·¥£Ê²Öͨ¨ ¢ · §²¨Î´ÒÌ Éμα Ì ±²¥Éμδμ£μ ͨ±² . ‚ ¶·μÍ¥¸¸¥ · ¡μÉÒ ¶μÖ¢¨²¨¸Ó ¤ ´´Ò¥ ¤·Ê£¨Ì ¢Éμ·μ¢ μ¡ ÊÎ ¸É¨¨ ±¨´ §Ò CDC28 ¢ Î¥±¶μ°´É-·¥£Ê²Öͨ¨ [32, 33], μ¤´ ±μ
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8215
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55,1±8,8
RAD17
rad17Δ
7
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7576
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43,1±7,8
RAD24
rad24Δ
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4462
4420
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rad53
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¢ ·¥£Ê²Öͨ¨ ±²¥Éμδμ£μ ͨ±² . ·μÉ¥¨´±¨´ § CDC28 μÉ´μ¸¨É¸Ö ± ¸¥³¥°¸É¢Ê
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¢Ò¸μ±μ¥ ¸·μ¤¸É¢μ ± ¤·μ¦¦¥¢μ° ±¨´ §¥ CDC28.
Š¨´ § CDC28 Ö¢²Ö¥É¸Ö ¶·¨¢²¥± É¥²Ó´μ° ³¨Ï¥´ÓÕ G1- ¨ G2-Î¥±¶μ°´Éμ¢,
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Y19 μ¡´ ·Ê¦¨¢ ¥É¸Ö ¢ Ë § Ì S ¨ G2, ´μ ´¥ μ¡´ ·Ê¦¨¢ ¥É¸Ö ¢ Ë § Ì G1 ¨ Œ.
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±É¨¢´μ¸ÉÓ ·34CDC28 ¢Ò¸μ± ¢ Ë §¥ S, ÌμÉÖ Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥ Y19 É ±¦¥
³ ±¸¨³ ²Ó´μ.
¸É ´μ¢± ±²¥Éμδμ£μ ͨ±² , ¢Ò§¢ ´´ Ö ´¥¤μ·¥¶²¨± ͨ¥° „Š, ¶μ-¢¨¤¨³μ³Ê, ´¥ ¸μ¶·μ¢μ¦¤ ¥É¸Ö ¸´¨¦¥´¨¥³ ±É¨¢´μ¸É¨ ·34CDC28 . ŒÊÉ Í¨¨, ¶·¥¤μÉ¢· Ð ÕШ¥ Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥ Y19 ±¨´ §Ò ·34CDC28 , ´¥ Ê¢¥²¨Î¨¢ ÕÉ
±¨´ §´ÊÕ ±É¨¢´μ¸ÉÓ Ê S- ·¥¸Éμ¢ ´´ÒÌ ±²¥Éμ± ¨, ´ μ¡μ·μÉ, ʸ²μ¢¨Ö, ¨´¤Êͨ·ÊÕШ¥ £¨¶¥·Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥ ·34CDC28 [43] ¨ ·¥¸É ¢ Ë §¥ G2 [44]
(¤¥²¥Í¨Ö MIH1 ¨ ¸¢¥·ÌÔ±¸¶·¥¸¸¨Ö wee1), ¶μ-¢¨¤¨³μ³Ê, ´¥ ¸´¨¦ ÕÉ ±¨´ §´μ°
±É¨¢´μ¸É¨, ± ± ¨ ¢ ¸²ÊÎ ¥ Ë §Ò S. “ ¤¥²¥Í¨μ´´μ£μ ³ÊÉ ´É ¶μ £¥´Ê MIH1
(MIH1 Ê S. cerevisiae £μ³μ²μ£¨Î¥´ cdc25 S. pombe) ´ ¡²Õ¤ ¥É¸Ö £¨¶¥·Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥ ’18 ¨ “19 ·34CDC28 . ¤´ ±μ § ³¥´ ɨ·μ§¨´ ´ Ë¥´¨² ² ´¨´
¨ É·¥μ´¨´ ´ ² ´¨´ (cdc28-[Y19F], cdc28-[T18A,Y19F]) ´¥ ¢Ò§Ò¢ ÕÉ § ³¥É´ÒÌ ¨§³¥´¥´¨° ¢·¥³¥´¨ £¥´¥· ͨ¨, · §³¥· ±²¥Éμ± ¨ · ¸¶·¥¤¥²¥´¨Ö ±²¥Éμ±
¶μ ¸É ¤¨Ö³ ±²¥Éμδμ£μ ͨ±² , μ¶·¥¤¥²Ö¥³ÒÌ ¸ ¶μ³μÐÓÕ FACS- ´ ²¨§ . μ²¥¥ Éμ£μ, 1-±¨´ § ´ Ö ±É¨¢´μ¸ÉÓ, ¸¢Ö§ ´´ Ö ¸ G2-¸¶¥Í¨Ë¨Î¥¸±¨³ ͨ±²¨´μ³
CLB2, ´¥ ¨§³¥´¥´ Ê ³ÊÉ ´É F19. ’ ±¨³ μ¡· §μ³, ¸É Éʸ Ëμ¸Ëμ·¨²¨·μ¢ ´¨Ö
21
Y19 ±¨´ §Ò ·34CDC28 ´¥ ¢ ¦¥´ ¤²Ö § ¢¨¸¨³μ¸É¨ ³¨Éμ§ μÉ § ¢¥·Ï¥´¨Ö ·¥¶²¨± ͨ¨ „Š. „²Ö G2-μ¸É ´μ¢±¨ ´¥ É·¥¡Ê¥É¸Ö É ±¦¥ ¸´¨¦¥´¨¥ ±É¨¢´μ¸É¨
±¨´ §Ò CDC28. ɨ ¤ ´´Ò¥ ¶·¥¤¶μ² £ ÕÉ, ÎÉμ Ê S. cerevisiae Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥ Y19 ³μ¦¥É ·¥£Ê²¨·μ¢ ÉÓ ¶·μÌ즤¥´¨¥ ±²¥Éμδμ£μ ͨ±² ³¥Ì ´¨§³μ³, ±μÉμ·Ò° ´¥ ¸¢μ¤¨É¸Ö ± ¶ ¤¥´¨Õ ± É ²¨É¨Î¥¸±μ° ±É¨¢´μ¸É¨ ·34CDC28 .
ŒÊÉ Í¨¨, ¶·¥¤μÉ¢· Ð ÕШ¥ Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥ ·34CDC28 ¶μ T18 ¨ Y19, ´¥
ʸ±μ·ÖÕÉ ³¨Éμ§ ¨ ´¥ ´ ·ÊÏ ÕÉ Î¥±¶μ°´É-·¥£Ê²ÖͨÕ, ¸¢Ö§Ò¢ ÕÐÊÕ ±²¥Éμδμ¥
¤¥²¥´¨¥ ¸ § ¢¥·Ï¥´¨¥³ ¸¨´É¥§ „Š.
„²Ö ¤¥²ÖÐ¨Ì¸Ö ¤·μ¦¦¥° S. pombe ¶μ± § ´μ, ÎÉμ μ¡· ¡μɱ ±²¥Éμ± HU
¢Ò§Ò¢ ¥É ¨´£¨¡¨·μ¢ ´¨¥ Ëμ¸Ëμ·¨²¨·μ¢ ´¨Ö ɨ·μ§¨´ Y15 ±¨´ §Ò Cdc2. ¤´ ±μ ³ÊÉ Í¨μ´´μ¥ ¨´£¨¡¨·μ¢ ´¨¥ ɨ·μ§¨´ Ê ¶μαÊÕÐ¨Ì¸Ö ¤·μ¦¦¥° ´¥ ¢²¨Ö²μ ´ S-Î¥±¶μ°´É, ¤¢μ°´ Ö ³ÊÉ Í¨Ö cdc28-[Y19F] rad53 ´¥ ʸ¨²¨¢ ¥É HU²¥É ²Ó´μ¸ÉÓ ³ÊÉ ´É rad53 [27]. ŒÊÉ ´É cdc28-[Y19F] ´¥ ¸¶μ¸μ¡¥´ ± ·¥¸ÉÊ ¢
μÉ¢¥É ´ ¤¥°¸É¢¨¥ HU ¨²¨ “”-μ¡²ÊÎ¥´¨¥ [45]. ɨ ³ÊÉ Í¨¨ ´¥ ¢²¨ÖÕÉ É ±¦¥
´ ÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ ± -²ÊÎ ³ ¨ HU [39, 43], ÌμÉÖ Ô±¢¨¢ ²¥´É´Ò° ³ÊÉ ´É S.
pombe ¢¸Éʶ ¥É ¢ ³¨Éμ§ ¶·¥¦¤¥¢·¥³¥´´μ [46].
’ ±¨³ μ¡· §μ³, ¢ μɲ¨Î¨¥ μÉ S. pombe, Xenopus ¨ ±²¥Éμ± ³²¥±μ¶¨É ÕШÌ
¤¥Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥ Y19 ´¥ É·¥¡Ê¥É¸Ö ¤²Ö ±É¨¢ ͨ¨ ±¨´ §Ò CDC28. μ²¥¥
Éμ£μ, ³ÊÉ Í¨μ´´Ò¥ § ³¥´Ò ÔÉμ£μ μ¸É ɱ ´¥ ¢²¨ÖÕÉ ´ § ¢¨¸¨³μ¸ÉÓ ³¨Éμ§ μÉ
¸¨´É¥§ „Š, ´¥ μɳ¥´ÖÕÉ G2- ·¥¸É, ¨´¤Êͨ·μ¢ ´´Ò° ¶μ¢·¥¦¤¥´¨Ö³¨ „Š.
μ-¢¨¤¨³μ³Ê, Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥ ɨ·μ§¨´ ´¥ Ö¢²Ö¥É¸Ö Ê´¨¢¥·¸ ²Ó´Ò³ ¨ ¸ÊÐ¥¸É¢Ê¥É ¤·Ê£μ° ±μ´É·μ²Ó ¢ÒÌμ¤ ¨§ ³¨Éμ§ . “ ³ÊÉ ´É cdc28-[Y19F] ±¨´ §´ Ö ±É¨¢´μ¸ÉÓ ³μ¦¥É ¨´£¨¡¨·μ¢ ÉÓ¸Ö Swe1p-§ ¢¨¸¨³Ò³ ¸¶μ¸μ¡μ³, ´¥§ ¢¨¸¨³Ò³ μÉ Ëμ¸Ëμ·¨²¨·μ¢ ´¨Ö ɨ·μ§¨´ [47]. ŒÊÉ Í¨Ö cdc28-[E12K] ¶·¨¢μ¤¨É
± ·¥§¨¸É¥´É´μ¸É¨ ± Swe1p-μ¶μ¸·¥¤μ¢ ´´μ³Ê ¨´£¨¡¨·μ¢ ´¨Õ. ŒÊÉ Í¨Ö cdc28[Y19F], ¶·¥¤μÉ¢· Ð ÕÐ Ö Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥ ±¨´ §Ò Cdc28 ±¨´ §μ° Swe1,
´¥ ¶·¥¤μÉ¢· Ð ¥É Swe1·-§ ¢¨¸¨³ÊÕ G2/M-§ ¤¥·¦±Ê, ¢Ò§¢ ´´ÊÕ £¨¶¥·Éμ´¨Î¥¸±¨³ Ïμ±μ³ [48].
“ ¤·μ¦¦¥° S. cerevisiae ¨ S. pombe ¸ÊÐ¥¸É¢ÊÕÉ ¢ ¦´Ò¥ · §²¨Î¨Ö ¢ ËÊ´±Í¨μ´¨·μ¢ ´¨¨ ³¨ÉμɨΥ¸±μ° Ëμ·³Ò ±¨´ §Ò CDC28/‘dc2. “ S. pombe ±É¨¢´μ¸ÉÓ ±¨´ §Ò ‘dc2 ¨³¥¥É ¶¨± Éμ²Ó±μ ¶¥·¥¤ ¢¸Éʶ²¥´¨¥³ ¢ Ë §Ê Œ [49, 50].
„ʶ²¨± ꬅ ¨ ³¨£· ꬅ ¶μ²Ö·´ÒÌ É¥²¥Í ³¨ÉμɨΥ¸±μ£μ ¢¥·¥É¥´ ¨ μ¡· §μ¢ ´¨¥ ³¨ÉμɨΥ¸±μ£μ ¢¥·¥É¥´ ¶·μ¨¸Ìμ¤¨É É ±¦¥ ¢ ÔÉμÉ ¶¥·¨μ¤ [51]. ¤´ ±μ
Ê ¶μαÊÕÐ¨Ì¸Ö ¤·μ¦¦¥° ¤Ê¶²¨± ꬅ ¶μ²Ö·´ÒÌ É¥²¥Í ¢¥·¥É¥´ ¨ ¨Ì ¸¥£·¥£ ꬅ ¶·μ¨¸Ìμ¤ÖÉ ³´μ£μ · ´ÓÏ¥ ¢ ±²¥ÉμÎ´μ³ Í¨±²¥, ¢¸±μ·¥ ¶μ¸²¥ § ¢¥·Ï¥´¨Ö
‘’’ [52, 53]. μÔÉμ³Ê ³ ²μ¢¥·μÖÉ´μ, ÎÉμ Ôɨ ¸μ¡ÒÉ¨Ö ¨´¨Í¨¨·ÊÕÉ¸Ö ±¨´ §μ°, § ¢¨¸¨³μ° μÉ Í¨±²¨´μ¢ CLB1 ¨ CLB2, ´μ ¡μ²¥¥ ¢¥·μÖÉ´μ, ÎÉμ Ôɨ
¸μ¡ÒÉ¨Ö ¨´¨Í¨¨·ÊÕÉ¸Ö ±¨´ §μ°, § ¢¨¸¨³μ° μÉ CLN1, CLN2, CLN3.
“ΨÉÒ¢ Ö ¸É·Ê±ÉÊ·´Ò¥ μ¸μ¡¥´´μ¸É¨ ³¨ÉμɨΥ¸±μ£μ ¶¶ · É ¶μαÊÕÐ¨Ì¸Ö ¤·μ¦¦¥°, ³μ¦´μ ¶·¥¤¶μ²μ¦¨ÉÓ, ÎÉμ ³¨Éμ§ ·¥£Ê²¨·Ê¥É¸Ö ¢ ¤¢ÊÌ Éμα Ì Å
¸¡μ·± ¢¥·¥É¥´ ±μ´É·μ²¨·Ê¥É¸Ö · ´ÓÏ¥, ±μ´É·μ²Ó ³¨ÉμɨΥ¸±μ° ¸¥£·¥£ ͨ¨
Ì·μ³μ¸μ³ μ¸ÊÐ¥¸É¢²Ö¥É¸Ö ¶μ§¦¥ [54]. ·¨ ÔÉμ³ · §²¨Î´Ò¥ ¸É ¤¨¨ ³¨Éμ§ 22
μ·±¥¸É·ÊÕÉ¸Ö ¤¢Ê³Ö ¨²¨ ¡μ²¥¥ Ëμ·³ ³¨ ±¨´ §Ò CDC28. ¤´ ¨§ ÔÉ¨Ì Ëμ·³,
¢μ§³μ¦´μ ¸μ¤¥·¦ Ð Ö CLB3 ¨ CLB4, ´¥μ¡Ì줨³ ¤²Ö ¨´¨Í¨ ͨ¨ ³¨Éμ§ ¨ μ¡· §μ¢ ´¨Ö ³¨ÉμɨΥ¸±μ£μ ¢¥·¥É¥´ , ¤·Ê£ Ö Ëμ·³ , ¸μ¤¥·¦ Ð Ö CLB1 ¨
CLB2, ´¥μ¡Ì줨³ ¤²Ö § ¢¥·Ï¥´¨Ö ³¨Éμ§ [55].
“ ¡μ²ÓϨ´¸É¢ ±²¥Éμ± ¤Ê¶²¨± ͨÖ, ¶·μ¨¸Ìμ¤ÖÐ Ö ¢ ¨´É¥·Ë §¥, ¸μ¶·μ¢μ¦¤ ¥É¸Ö Ê¢¥²¨Î¥´¨¥³ · §³¥· ±²¥É±¨. ¤´ ±μ Ê S. cerevisiae ¤μÎ¥·´ÖÖ ±²¥É± (¶μα ) ¸É·μ¨É¸Ö de novo μɤ¥²Ó´μ μÉ ³ É¥·¨´¸±μ° ±²¥É±¨. ɨ ¶·μÍ¥¸¸Ò
É·¥¡ÊÕÉ ¸¥·¨¨ ¨§³¥´¥´¨° ¢ ¶μ²Ö·´μ¸É¨ ·μ¸É ¤²Ö ¸μ§·¥¢ ´¨Ö ¶μα¨ ´ ¶μ¢¥·Ì´μ¸É¨ ³ É¥·¨´¸±μ° ±²¥É±¨. μ± § ´μ, ÎÉμ ±É¨¢ ꬅ ±μ³¶²¥±¸μ¢ CLB1¨ CLB2-CDC28 É·¥¡Ê¥É¸Ö ¤²Ö ¢±²ÕÎ¥´¨Ö ¤¥¶μ²Ö·¨§μ¢ ´´μ£μ ·μ¸É ¶μα¨ ¢
Ë §¥ G2 [56]. Š¨´ §´ Ö ±É¨¢´μ¸ÉÓ CLB2-CDC28 ´¥μ¡Ì줨³ ¤²Ö ¨´¨Í¨ ͨ¨
¶μ²μ¦¥´¨Ö ¶μα¨ [57] ¨ ¶¥·¥±²ÕÎ¥´¨Ö μÉ ¢¥·ÌÊϥδμ£μ ± ¨§μÉ·μ¶´μ³Ê ·μ¸ÉÊ
¶μα¨ [58]. …¸²¨ Ôɨ Ëμ·³Ò ±¨´ §Ò ¨´£¨¡¨·μ¢ ´Ò (Ô²¨³¨´ ͨ¥° ͨ±²¨´μ¢
¨²¨ ¨´£¨¡¨Éμ·´Ò³ Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥³ ɨ·μ§¨´ ), ¶μα¨ ¸É ´μ¢ÖÉ¸Ö ´¥´μ·³ ²Ó´μ ¢ÒÉÖ´ÊÉÒ³¨. …¸²¨ Î¥±¶μ°´É μ¸ÊÐ¥¸É¢²Ö¥É¸Ö É¥³ ¦¥ ³¥Ì ´¨§³μ³, ÎÉμ
¨ ¢ ±²¥É± Ì S. pombe ¨²¨ Xenopus, Éμ ±²¥É±¨ ¸ ´¥´μ·³ ²Ó´μ° ¶μ¢¥·Ì´μ¸ÉÓÕ ¡Ê¤ÊÉ μ¡· §μ¢Ò¢ ÉÓ¸Ö ¨ ¢ ʸ²μ¢¨ÖÌ, ±μ£¤ ±²¥É±¨ 즨¤ ÕÉ § ¢¥·Ï¥´¨Ö
·¥¶²¨± ͨ¨ ¨/¨²¨ ·¥¶ · ͨ¨. ‚ ¸²ÊÎ ¥ μ¡· ¡μɱ¨ ±²¥Éμ± HU ´ ¡²Õ¤ ¥É¸Ö
¨´ Ö ± ·É¨´ : CLB-CDC28-μ¶μ¸·¥¤μ¢ ´´μ¥ ¸μ§·¥¢ ´¨¥ ¶μα¨ ¶·μ¨¸Ìμ¤¨É ± ±
μ¡ÒÎ´μ ¨ ¶ · ³ É¥·¨´¸± ÖĤμÎ¥·´ÖÖ ±²¥É±¨ · ¸ÉÊÉ ¢ ¶·μÍ¥¸¸¥ ·¥¸É . ‚μ§³μ¦´μ, ³¥Ì ´¨§³ Î¥±¶μ°´É-·¥£Ê²Öͨ¨ Ê S. cerevisiae Ô¢μ²ÕÍ¨μ´¨·μ¢ ² ¢ ¸Éμ·μ´Ê § ¢¥·Ï¥´¨Ö CLB-CDC28-μ¶μ¸·¥¤μ¢ ´´ÒÌ ³μ·Ëμ£¥´¥É¨Î¥¸±¨Ì ¸μ¡Òɨ°,
ÌμÉÖ ¶·¨ ÔÉμ³ ¡²μ±¨·μ¢ ´Ò Ö¤¥·´Ò¥ ¸μ¡ÒɨÖ.
μ± § ´μ, ÎÉμ ¤¨¶²μ¨¤, £μ³μ§¨£μÉ´Ò° ¶μ ³ÊÉ Í¨¨ cdc28-[T18A,Y19F],
¶·μÖ¢²Ö¥É ¤¥Ë¥±É´μ¸ÉÓ ¶μ ˨² ³¥´É´μ³Ê ·μ¸ÉÊ ¨ ¸´¨¦¥´´ÊÕ Ê¤²¨´¥´´μ¸ÉÓ
±²¥É±¨ [59]. ŒÊÉ ´É cdc28-[Y19E] ´¥ ¸¶μ¸μ¡¥´ ± ¸¥£·¥£ ͨ¨ ¤Ê¶²¨Í¨·μ¢ ´´ÒÌ ¶μ²Ö·´ÒÌ É¥²¥Í ¢¥·¥É¥´ [60].
μ²ÊÎ¥´´ Ö ´ ³¨ ³ÊÉ Í¨Ö cdc28-srm-[G20S] ¢²¨Ö¥É ´ Ëμ·³Ê ±²¥Éμ± ¨
´ ³μ¤¥²Ó ¶μÎ±μ¢ ´¨Ö [1]. Š·μ³¥ Éμ£μ, ± ± ¶μ± § ´μ ¢ ´ ¸ÉμÖÐ¥° · ¡μÉ¥,
μ´ ¢²¨Ö¥É ¶· ±É¨Î¥¸±¨ ´ ¢¸¥ Î¥±¶μ°´ÉÒ. Š ¸μ¦ ²¥´¨Õ, ´¥ ¨§¢¥¸É´μ ¶·¨¢μ¤¨É ²¨ ÔÉ ³ÊÉ Í¨Ö ± ¨§³¥´¥´¨Õ ±¨´ §´μ° ±É¨¢´μ¸É¨ ± ±μ°-²¨¡μ Ëμ·³Ò.
¤´ ±μ ·¥§Ê²ÓÉ ÉÒ Œ„-³μ¤¥²¨·μ¢ ´¨Ö ¶μ± § ²¨, ÎÉμ § ³¥´ G20S ¢Ò§Ò¢ ¥É
¸¨²Ó´Ò¥ ¨§³¥´¥´¨Ö ±μ´Ëμ·³ ͨ¨ ¸ ³μ° ±¨´ §Ò, ¥¥ ±μ³¶²¥±¸μ¢ ¸ ͨ±²¨´ ³¨
¨ ¸Ê¡¸É· Éμ³, ¶μ-¢¨¤¨³μ³Ê, ³¥´ÖÖ ¸Ê¡¸É· É´ÊÕ ¸¶¥Í¨Ë¨Î´μ¸ÉÓ ±¨´ §Ò [61].
´¥¥ ¡Ò² μ¡´ ·Ê¦¥´ ³ÊÉ Í¨Ö cdc28-5M, ¶·¥¤¸É ¢²ÖÕÐ Ö ¸μ¡μ° ³´μ¦¥¸É¢¥´´Ò¥ § ³¥´Ò (L11H, L183S, S216C, E217K, N232D), ±μÉμ· Ö É ±¦¥
´ ·ÊÏ ² μ¸É ´μ¢±Ê ±²¥Éμδμ£μ ͨ±² , ¨´¤Êͨ·μ¢ ´´ÊÕ “”-¶μ¢·¥¦¤¥´¨Ö³¨
¨ ´μ±μ¤ §μ²μ³, ´μ ´¥ £¨¤·μ±¸¨³μÎ¥¢¨´μ° HU [32]. ɳ¥É¨³, ÎÉμ 줨´μδҥ ³ÊÉ Í¨¨, ¢ Éμ³ Î¨¸²¥ [L11H] (²μ± ²¨§μ¢ ´´ Ö ¢ Éμ° ¦¥ G-¡μ£ Éμ° ¶¥É²¥,
ÎÉμ ¨ ³ÊÉ Í¨Ö cdc28-srm-[G20S]), ´¥ ¨³¥ÕÉ ¢¨¤¨³ÒÌ ¶·μÖ¢²¥´¨°. “ ³ÊÉ ´É cdc-5M ´ ¡²Õ¤ ¥É¸Ö ¸´¨¦¥´¨¥ ±¨´ §´μ° ±É¨¢´μ¸É¨, ¢±²ÕÎ Ö CLB2-, CLB5-,
CLB6-, ´μ ´¥ CLB3- ¸¸μͨ¨·μ¢ ´´μ° CDK- ±É¨¢´μ¸É¨. –¨±²¨´Ò CLB5 ¨
23
CLB6 Ô±¸¶·¥¸¸¨·ÊÕÉ¸Ö ¢ ‘’’¥ ¨ ´¥μ¡Ì줨³Ò ¤²Ö ¨´¨Í¨ ͨ¨ S, ͨ±²¨´ CLB2 Ô±¸¶·¥¸¸¨·Ê¥É¸Ö ¢ G2/M ¨ ´¥μ¡Ì줨³ ¤²Ö ³¨Éμ§ . –¨±²¨´ CLB3
Ô±¸¶·¥¸¸¨·ÊÉ¸Ö ¢ S/G2 ¨ ¢ ¦¥´ ¤²Ö μ¡· §μ¢ ´¨Ö ¢¥·¥É¥´ . ’ ±¨³ μ¡· §μ³, Ê
³ÊÉ ´É ´ ·ÊÏ¥´ ±¨´ §´ Ö ±É¨¢´μ¸ÉÓ ¢ ‘’’¥ ¨ ³¨É짥, ´μ ´¥ ¢ S/G2.
‘² ¡Ò° ÔËË¥±É ´ ËÊ´±Í¨μ´¨·μ¢ ´¨¥ Î¥±¶μ°´É μ¡´ ·Ê¦¥´ Ê ³ÊÉ ´É´ÒÌ ²²¥²¥° cdc28-9 ¨ cdc28-13. “ ³ÊÉ ´Éμ¢ cdc28-4, -9, -13, -5M [32], É ± ¦¥ ± ±
¨ Ê ³ÊÉ ´É cdc28-srm [1], ¸´¨¦¥´ ¸¶μ¸μ¡´μ¸ÉÓ ¶μ¤¤¥·¦ ´¨Ö Í¥´É·μ³¥·´μ°
¶² §³¨¤Ò.
Œ¨Ï¥´ÓÕ ·¥£Ê²Öͨ¨ ¶μ³¨³μ ± É ²¨É¨Î¥¸±μ° ¸Ê¡Ñ¥¤¨´¨ÍÒ ³μ£ÊÉ ¸²Ê¦¨ÉÓ
ͨ±²¨´Ò. ˆ§¢¥¸É´μ, ÎÉμ ¸¢¥·ÌÔ±¸¶·¥¸¸¨Ö ͨ±²¨´μ¢ μ¸² ¡²Ö¥É ¶·μÖ¢²¥´¨Ö
¤¥Ë¥±Éμ¢ Î¥±¶μ°´É [62]. “ ¤·μ¦¦¥° ¶μ¢·¥¦¤¥´¨Ö „Š ±É¨¢¨·ÊÕÉ ±¨´ §Ê Rad53, ±μÉμ· Ö ¨´ ±É¨¢¨·Ê¥É É· ´¸±·¨¶Í¨μ´´Ò° ·¥£Ê²ÖÉμ· Swi6, ¨´£¨¡¨·ÊÕШ° É· ´¸±·¨¶Í¨μ´´Ò° ±É¨¢ Éμ· Swi4 É· ´¸±·¨¶Í¨¨ G1-ͨ±²¨´μ¢
(£¥´Ò CLN). ’ ±¨³ μ¡· §μ³, CDK-ͨ±²¨´Ò, ¢ ´μ·³¥ ´¥μ¡Ì줨³Ò¥ ¤²Ö G1/S¶¥·¥Ìμ¤ , ¨´ ±É¨¢¨·ÊÕÉ¸Ö ¶μ¸²¥ ¶μ¢·¥¦¤¥´¨Ö ³μ²¥±Ê²Ò „Š. “ ¤·μ¦¦¥°
μ¸É ´μ¢± ¢ G1/S ¨ G2/M, ¶μ-¢¨¤¨³μ³Ê, μ¡¥¸¶¥Î¨¢ ¥É ¢·¥³Ö ¤²Ö ·¥¶ · ͨ¨.
“ ³²¥±μ¶¨É ÕÐ¨Ì μ¸É ´μ¢± ¢ G1/S ´Ê¦´ ´¥ ¤²Ö ·¥¶ · ͨ¨, ¸±μ·¥¥ ¤²Ö
¢±²ÕÎ¥´¨Ö ²ÓÉ¥·´ ɨ¢´μ£μ ¶Êɨ, ¶·¨¢μ¤ÖÐ¥£μ ± £¨¡¥²¨ ±²¥É±¨ ( ¶μ¶Éμ§Ê).
Š·μ³¥ Éμ£μ, ±É¨¢´μ¸ÉÓ ±¨´ §Ò ³μ¦¥É ³¥´ÖÉÓ¸Ö É¨·μ§¨´-´¥§ ¢¨¸¨³Ò³
¸¶μ¸μ¡μ³, ´ ¶·¨³¥· § ¸Î¥É ¸¨´É¥§ ¨´£¨¡¨Éμ· CDK Å Sic1. ‚¥·μÖÉ´μ° ³¨Ï¥´ÓÕ Ö¢²Ö¥É¸Ö É ±¦¥ ¡¥²μ± Cks1 [63], £μ³μ²μ£ suc1+ S. pombe.
·13suc1 , ¸¢Ö§Ò¢ Ö¸Ó ¸ ‘DC28/ͨ±²¨´μ¢Ò³¨ ±μ³¶²¥±¸ ³¨, ³μ¦¥É ³¥´ÖÉÓ ¸Ê¡¸É· É´ÊÕ ¸¶¥Í¨Ë¨Î´μ¸ÉÓ ±¨´ §Ò [64]. •μÉÖ ¡¨μ̨³¨Î¥¸±μ¥ §´ Î¥´¨¥ ¸¢Ö§Ò¢ ´¨Ö ¡¥²±μ¢ ·18CKS1 ¨ ·34CDC28 ´¥¨§¢¥¸É´μ, ¡¥²μ± Cks1 ´¥μ¡Ì줨³ ¤²Ö
¶¥·¥Ìμ¤μ¢ G1/S ¨ G2/M [65] ¨ μ¡´ ·Ê¦¨¢ ¥É¸Ö ¢ ¸¸μͨ ͨ¨ ¸ CDC28 ¢
HU-μ¡· ¡μÉ ´´ÒÌ ±²¥É± Ì [62]. μ¸±μ²Ó±Ê ³ÊÉ Í¨Ö cks1-ts38 ¸Ê¶·¥¸¸¨·Ê¥É
HU-ÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ ³ÊÉ ´É rad53, ¶μ-¢¨¤¨³μ³Ê, ¡¥²μ± Cks1 μ¸ÊÐ¥¸É¢²Ö¥É
´¥£ ɨ¢´ÊÕ ·¥£Ê²ÖÍ¨Õ ¢ RAD53-§ ¢¨¸¨³μ³ ¨´£¨¡¨·μ¢ ´¨¨ ¶·μÌ즤¥´¨Ö ±²¥Éμδμ£μ ͨ±² [27].
ˆ¸¶μ²Ó§μ¢ ´¨¥ ³ÊÉ Í¨¨ cdc28-as1 [F88G] ¶μ± § ²μ, ÎÉμ ±¨´ § CDC28
´¥μ¡Ì줨³ ¤²Ö ±É¨¢ ͨ¨ „ „Š Œ¥¸1·-§ ¢¨¸¨³μ° μ¸É ´μ¢±¨ ±²¥Éμδμ£μ
ͨ±² ¢ Ë §¥ G2 [33]. Š ¸Ê¡¸É· É ³ CDK μÉ´μ¸ÖÉ¸Ö É ±¨¥ Î¥±¶μ°´É-¡¥²±¨,
± ± Rad9, Drc1/Sld2, Ddc1, Dun1 ¨ Ptc3 [66Ä68]. „²Ö ±É¨¢ ͨ¨ Î¥±¶μ°´É±μ´É·μ²Ö ´¥μ¡Ì줨³Ò ¤¥£· ¤ ꬅ ±μ´Íμ¢ ¤¢Ê´¨É¥¢μ£μ · §·Ò¢ „Š, § £·Ê§± RPA ´ ÊÎ ¸Éμ± μ¤´μ´¨É¥¢μ° „Š ¨ ¶μ¸²¥¤ÊÕÐ Ö § £·Ê§± ±¨´ §Ò
Mec1 [69, 70]. ˆ´£¨¡¨·μ¢ ´¨¥ ±¨´ §´μ° ±É¨¢´μ¸É¨ CDC28 ´ ·ÊÏ ¥É ±É¨¢ Í¨Õ μ¸É ´μ¢±¨, ¨´¤Êͨ·μ¢ ´´μ° ¤¢Ê´¨É¥¢Ò³ · §·Ò¢μ³ „Š, ´μ ´¥ ¸ÊÐ¥¸É¢¥´´μ ¤²Ö ¤·Ê£¨Ì ¶μ¢·¥¦¤¥´¨°, ¨´¤Êͨ·ÊÕÐ¨Ì Œ¥¸1·-§ ¢¨¸¨³ÊÕ ±É¨¢ Í¨Õ Î¥±¶μ°´É-±μ´É·μ²Ö. ·¥¤¶μ² £ ¥É¸Ö, ÎÉμ CDC28 ¢²¨Ö¥É ´ ±É¨¢ ͨÕ
Œ¥¸1·-§ ¢¨¸¨³μ° μ¸É ´μ¢±¨ Éμ²Ó±μ ¢ ¸²ÊÎ ¥, ¥¸²¨ ¶μ¢·¥¦¤¥´¨¥ ¶·¥¤¸É ¢²Ö¥É
¸μ¡μ° ¤¢Ê´¨É¥¢μ° · §·Ò¢ „Š, ´Ê¦¤ ÕШ°¸Ö ¢ ¤¥£· ¤ ͨ¨ ¤²Ö μ¡· §μ¢ ´¨Ö
μ¤´μ´¨É¥¢μ° „Š.
24
쳨³μ ±É¨¢ ͨ¨ Î¥±¶μ°´É ±¨´ § CDC28 ÊÎ ¸É¢Ê¥É ´ ˨´ ²Ó´ÒÌ ÔÉ ¶ Ì ·¥£Ê²Öͨ¨ μ¸É ´μ¢±¨ ±²¥Éμδμ£μ ͨ±² . „²Ö ¢ÒÌμ¤ ¨§ ³¨Éμ§ ¨ § ¢¥·Ï¥´¨Ö ±²¥Éμδμ£μ ͨ±² É·¥¡Ê¥É¸Ö ¨´ ±É¨¢ ꬅ ±¨´ §Ò CDC28/CLB2. ·μÉ¥ ¸μ³ APCCdc20 μ¶μ¸·¥¤Ê¥É Ê¡¨±¢¨É¨´¨§ Í¨Õ ¨ · §·ÊÏ¥´¨¥ ´ Ë §´μ£μ ¨´£¨¡¨Éμ· Pds1, É ±¦¥ ·¥£Ê²¨·Ê¥É ¶¥·¢Ò° Ê·μ¢¥´Ó · §·ÊÏ¥´¨Ö ͨ±²¨´ CLB2,
¶·¨¢μ¤ÖШ° ± ±É¨¢ ͨ¨ ¸¥É¨ Œ…N (mitotic exit network), ±μÉμ· Ö ·¥£Ê²¨·Ê¥É
¢ÒÌμ¤ ¨§ ³¨Éμ§ . ±É¨¢ ꬅ APCHct1/Cdh1 ¨ ¸¨´É¥§ ¨´£¨¡¨Éμ· CDC28 (Sic1)
¶μ§¢μ²ÖÕÉ μ¸ÊÐ¥¸É¢¨ÉÓ ¤ ²Ó´¥°ÏÊÕ ¨´ ±É¨¢ Í¨Õ ±μ³¶²¥±¸ CDC28/CLB2.
·¨ § ¤¥·¦±¥ ±²¥Éμδμ£μ ͨ±² ¢ Ë §¥ G2/M ±¨´ § RAD53 ¶μ¤¤¥·¦¨¢ ¥É
¢Ò¸μ±ÊÕ ±É¨¢´μ¸ÉÓ ±¨´ §Ò CDC28 Î¥·¥§ ¨´£¨¡¨·μ¢ ´¨¥ ±¨´ §Ò CDC5, ±μÉμ· Ö ´¥μ¡Ì줨³ ¤²Ö ¤¥£· ¤ ͨ¨ ͨ±²¨´μ¢ ¨ ¢ÒÌμ¤ ¨§ ³¨Éμ§ [71]. Š¨´ § RAD53 ÊÎ ¸É¢Ê¥É ¢ ±¨´ §´μ³ ± ¸± ¤¥ ¶¥·¥¤ Ψ ¸¨£´ ² ¶μ¸·¥¤¸É¢μ³ ±¨´ §Ò
DUN1, ¨´£¨¡¨·ÊÖ ¤¥£· ¤ Í¨Õ CLB2 ¨ ¢ÒÌμ¤ ¨§ ³¨Éμ§ , ¡²μ±¨·ÊÖ ±É¨¢ ͨÕ
Œ…N-±μ³¶²¥±¸ . ¤´ ±μ ¢ ´ Ï¨Ì Ô±¸¶¥·¨³¥´É Ì Ê ³ÊÉ ´É cdc28-srm ´¥ ¢ÒÖ¢²¥´μ ´ ·ÊÏ¥´¨¥ G2-Î¥±¶μ°´É , ¨´¤Êͨ·μ¢ ´´μ£μ γ-¨§²ÊÎ¥´¨¥³.
±É¨¢´μ¸ÉÓ CDC28 ´¥ ´Ê¦´ ¤²Ö ±É¨¢ ͨ¨ Î¥±¶μ°´É ¶·¥¤¸ÊÐ¥¸É¢ÊÕШ³¨ ¨´É¥·³¥¤¨ É ³¨ μ¤´μ´¨É¥¢μ° „Š ¢ ¶·μÍ¥¸¸¥ Ô±¸Í¨§¨μ´´μ° ·¥¶ · ͨ¨
´Ê±²¥μɨ¤μ¢. ¤´ ±μ ³Ò ´ ¡²Õ¤ ²¨ ¸´¨¦¥´¨¥ § ¤¥·¦±¨ ¶·¨ “”-μ¡²ÊÎ¥´¨¨
±Ê²ÓÉÊ·, ¸¨´Ì·μ´¨§μ¢ ´´ÒÌ ¢ Ë § Ì G0 ¨²¨ G1. μ ´ Ϩ³ ¤ ´´Ò³, ³ÊÉ Í¨Ö cdc28-srm ´¥ ¢²¨Ö¥É ´ S- ·¥¸É ¶μ¤ ¤¥°¸É¢¨¥³ HU, ´μ ¢²¨Ö¥É ´ intra
S-Î¥±¶μ°´É. ‚ ¤·Ê£μ° · ¡μÉ¥ ¶μ± § ´μ, ÎÉμ ¨´£¨¡¨·μ¢ ´¨¥ ±¨´ §´μ° ±É¨¢´μ¸É¨ CDC28 ´¥ ¢²¨Ö¥É ´ ±É¨¢ Í¨Õ Î¥±¶μ°´É ¢ μÉ¢¥É ´ ·¥¸É S-Ë §Ò,
¢Ò§¢ ´´Ò° HU, ¢ ¶·μÍ¥¸¸¥ ±μÉμ·μ£μ ¢μ§´¨± ÕÉ ÊÎ ¸É±¨ μ¤´μ´¨É¥¢μ° „Š
¢ § ¸É·Ö¢Ï¨Ì ·¥¶²¨± ɨ¢´ÒÌ ¢¨²± Ì [72]. ’ ±¨³ μ¡· §μ³, ±¨´ § CDC28,
ÊÎ ¸É¢ÊÕÐ Ö ¢ ±É¨¢ ͨ¨ ¸¨´É¥§ „Š ¨ ¶¥·¥Ì줥 ¨§ Ë §Ò G1 ¢ Ë §Ê S,
ÊÎ ¸É¢Ê¥É ¢ G1/S-Î¥±¶μ°´É¥, ´μ ´¥ ´Ê¦´ ´ ¸É ¤¨¨, ±μ£¤ ·¥¶²¨± Í¨Ö Ê¦¥
´ Î ² ¸Ó, ´ ¶·¨³¥·, ¶·¨ ¨´£¨¡¨·μ¢ ´¨¨ ·¥¶²¨± ͨ¨ HU.
’ ±¨³ μ¡· §μ³, ±¨´ § CDC28 ¶·¨´¨³ ¥É ±É¨¢´μ¥ ÊÎ ¸É¨¥ ¢ ¶·μÌ즤¥´¨¨ ±²¥Éμδμ£μ ͨ±² , μ¡¥¸¶¥Î¨¢ Ö ±É¨¢ Í¨Õ É· ´¸±·¨¶Í¨¨ ¸μμÉ¢¥É¸É¢ÊÕÐ¨Ì £¥´μ¢ ¨²¨ μ¸ÊÐ¥¸É¢²ÖÖ ±É¨¢ Í¨μ´´μ¥ Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥ ¡¥²±μ¢, ¨
¸²Ê¦¨É ³¨Ï¥´ÓÕ Î¥±¶μ°´É-·¥£Ê²Öͨ¨. Š·μ³¥ Éμ£μ, μ´ ¶·¨´¨³ ¥É ÊÎ ¸É¨¥ ´ ´ Î ²Ó´ÒÌ ÔÉ ¶ Ì ±É¨¢ ͨ¨ μ¸É ´μ¢±¨, Ëμ¸Ëμ·¨²¨·ÊÖ ¡¥²±¨, ÊÎ ¸É¢ÊÕШ¥
¢ ·¥¶ · ͨ¨, ·¥±μ³¡¨´ ͨ¨ ¨ Î¥±¶μ°´É¥. ±É¨¢´μ¸ÉÓ ±¨´ §Ò, ¶μ-¢¨¤¨³μ³Ê,
·¥£Ê²¨·Ê¥É¸Ö ´¥ Éμ²Ó±μ ¸É ÉÊ¸μ³ Ëμ¸Ëμ·¨²¨·μ¢ ´¨Ö ± É ²¨É¨Î¥¸±μ° ¸Ê¡Ñ¥¤¨´¨ÍÒ, ´μ ¨ ´ Ê·μ¢´¥ ¸¨´É¥§ ͨ±²¨´μ¢ ¨/¨²¨ ¨´£¨¡¨Éμ· .
³¨ ¢¶¥·¢Ò¥ ¶μ± § ´μ ÊÎ ¸É¨¥ ¢ Î¥±¶μ°´É-·¥£Ê²Öͨ¨ ±·μ³¥ ±¨´ §Ò
CDC28 £¥´μ¢ NET1 ¨ HFI1. HFI1/SRM12/ADA1 ¢Ìμ¤¨É ¢ ¸μ¸É ¢ ±μ ±É¨¢ Éμ·μ¢ É· ´¸±·¨¶Í¨¨ SAGA ¨ GCN5/ADA, μ¡² ¤ ÕÐ¨Ì £¨¸Éμ´ Í¥É¨²É· ´¸Ë¥· §´μ° ±É¨¢´μ¸ÉÓÕ HAT (histone acetyltransferase) [73]. ¤´ ±μ ͥɨ² § Gcn5, ¢Ìμ¤ÖÐ Ö ¢ ¸μ¸É ¢ ÔÉ¨Ì ±μ³¶²¥±¸μ¢, ËÊ´±Í¨μ´¨·Ê¥É ´¥ Éμ²Ó±μ ± ± ±μ ±É¨¢ Éμ· É· ´¸±·¨¶Í¨¨, ´μ ¨ μ¸ÊÐ¥¸É¢²Ö¥É ¥Ð¥ £²μ¡ ²Ó´μ¥ ͥɨ²¨·μ¢ ´¨¥
£¨¸Éμ´μ¢ ¶μ ¢¸¥³Ê £¥´μ³Ê [74]. ‚ ¶μ¸²¥¤´¥¥ ¢·¥³Ö ¸É ´μ¢¨É¸Ö μÎ¥¢¨¤´Ò³, ÎÉμ
25
ͥɨ²¨·μ¢ ´¨¥/¤¥ ͥɨ²¨·μ¢ ´¨¥ £¨¸Éμ´μ¢ ¨£· ¥É ·¥£Ê²ÖÉμ·´ÊÕ ·μ²Ó ¢ ·¥¶ · ͨ¨ ¨ ¢ ±μ´É·μ²¥ Î¥±¶μ°´É-μ¸É ´μ¢±¨. ¶·¨³¥·, ÊÉ· É Í¥É¨²¨·μ¢ ´´ÒÌ
²¨§¨´μ¢ Lys5, 8, 12 ¨ 16 £¨¸Éμ´ H4 ¨²¨ ´ ·ÊÏ¥´¨¥ Esa1p, ±μ³¶μ´¥´É HAT-±μ³¶²¥±¸ NuA4 ±É¨¢¨·ÊÕÉ RAD9-§ ¢¨¸¨³Ò° G2/M-Î¥±¶μ°´É [75Ä77].
§·ÊÏ¥´¨¥ £¨¸Éμ´¤¥ ͥɨ²É· ´¸Ë¥· §Ò HDAC (histone deacetyltransferase)
Sin3/Rpd3 ¢μ¸¸É ´ ¢²¨¢ ¥É G2/M- ·¥¸É ±²¥Éμδμ£μ ͨ±² ¶μ¸²¥ “”-μ¡²ÊÎ¥´¨Ö Ê Î¥±¶μ°´É-¤¥Ë¥±É´ÒÌ ÏÉ ³³μ¢ [78], ¨ ÔÉμ ¢μ¸¸É ´μ¢²¥´¨¥ § ¢¨¸¨É μÉ
¨´É ±É´μ£μ MAD1-§ ¢¨¸¨³μ£μ Î¥±¶μ°´É-±μ´É·μ²Ö ¢¥·¥É¥´ . ƒ¨¸Éμ´¤¥ ͥɨ²É· ´¸Ë¥· § Sin3/Rpd3 É ±¦¥ ·¥£Ê²¨·Ê¥É ¢·¥³Ö ·¥¶²¨± ͨ¨ ¶μ§¤´¨Ì ori ¨
ÊÎ ¸É¢Ê¥É ¢ intra S-Î¥±¶μ°´É¥ [79]. ˆ§¢¥¸É´Ò ¤ ´´Ò¥ μ¡ ÊÎ ¸É¨¨ ͥɨ²¨·μ¢ ´¨Ö/¤¥ ͥɨ²¨·μ¢ ´¨Ö ´¥£¨¸Éμ´μ¢ÒÌ ¡¥²±μ¢ ¢ ±μ´É·μ²¥ ¶μ¶Éμ§ ¨ · ¤¨μÎÊ¢¸É¢¨É¥²Ó´μ¸É¨ ±²¥Éμ± Î¥²μ¢¥± [80Ä84]. ¥²μ± hADA3, ¢Ìμ¤ÖШ° ¢ ¸μ¸É ¢
HAT-±μ³¶²¥±¸μ¢, Ö¢²Ö¥É¸Ö ±μË ±Éμ·μ³ ±É¨¢ ͨ¨ É· ´¸±·¨¶Í¨μ´´μ£μ Ë ±Éμ· p53 [85], ÊÎ ¸É¢ÊÕÐ¥£μ ¢ G1- ·¥¸É¥ ¨ ¶μ¶É짥. ͥɨ²¨·μ¢ ´¨¥ ·53
ͥɨ²É· ´¸Ë¥· §μ° PCAF/yGCN5 ·¥£Ê²¨·Ê¥É ±É¨¢´μ¸ÉÓ p53 ¢ ¶·μÍ¥¸¸¥ μÉ¢¥É ´ ¶μ¢·¥¦¤¥´¨Ö „Š [80-82, 86, 87]. p53-μ¶μ¸·¥¤μ¢ ´´Ò° ·¥£Ê²ÖÉμ·´Ò°
¶ÊÉÓ É ±¦¥ μ¶μ¸·¥¤Ê¥É¸Ö NAD-§ ¢¨¸¨³Ò³ ¤¥ ͥɨ²¨·μ¢ ´¨¥³ £¨¸Éμ´μ¢ ¤¥ ͥɨ² §μ° Sir2α [83, 84].
ˆ§ÊÎ¥´¨¥ ·μ²¨ ¶·μ¤Ê±Éμ¢ £¥´μ¢ CDC28, NET1 ¨ HFI1 ¢ Î¥±¶μ°´É-·¥£Ê²Öͨ¨ ¢Ò§Ò¢ ¥É ´¥¸μ³´¥´´Ò° ¨´É¥·¥¸ ¨ É·¥¡Ê¥É ¤ ²Ó´¥°Ï¨Ì ¨¸¸²¥¤μ¢ ´¨°.
·¥¤¸É ¢²Ö¥É ¨´É¥·¥¸ ¨§ÊÎ¥´¨¥ ¢§ ¨³μ¤¥°¸É¢¨Ö ³¥¦¤Ê ¶·μ¤Ê±É ³¨ ÔÉ¨Ì £¥´μ¢. ¥¤ ¢´μ μ¡´ ·Ê¦¨²¨ ¸¢Ö§Ó ³¥¦¤Ê ±¨´ §μ° CDC28 ¨ ’- ±É¨¢´μ¸ÉÓÕ,
¨³¥´´μ, ¶μ± § ´μ, ÎÉμ Ëμ¸Ëμ·¨²¨·μ¢ ´¨¥ £¨¸Éμ´ 3 ¶μ ¸¥·¨´Ê Ser10 ±¨´ §μ° Snf1 μ¡²¥£Î ¥É ¤ ²Ó´¥°Ï¥¥ ͥɨ²¨·μ¢ ´¨¥ ²¨§¨´ Lys14 ͥɨ² §μ°
Gcn5 [88, 89]. ¥£ ɨ¢´Ò° ·¥£Ê²ÖÉμ· Sip2 ±¨´ §Ò Snf1 (¸Ê¡Ñ¥¤¨´¨Í ±μ³¶²¥±¸ SWI/SNF) Ö¢²Ö¥É¸Ö ¸Ê¡¸É· Éμ³ ¶·μÉ¥¨´±¨´ §Ò CDC28 [66]. Š·μ³¥
Éμ£μ, ¸Ê¡¸É· É ³¨ ±¨´ §Ò CDC28 Ö¢²ÖÕÉ¸Ö ¡¥²±¨ É· ´¸±·¨¶Í¨μ´´ÒÌ ±μ³¶²¥±¸μ¢ TFG1 (TFIIF), TAF4 (TFIID), NGG1 (SAGA), SPT16 [66], ¶·¨Î¥³
SPT16 ¸¢Ö§Ò¢ ¥É¸Ö ¸ ’-±μ³¶²¥±¸μ³ NuA3 [90].
¢Éμ·Ò ¶·¨´μ¸ÖÉ ¡² £μ¤ ·´μ¸ÉÓ ’. . §²μ¢μ° § ɥ̴¨Î¥¸±ÊÕ ¶μ³μÐÓ
¨ . . „¥¢¨´Ê § ¸±μ´¸É·Ê¨·μ¢ ´´Ò¥ ÏÉ ³³Ò ¨ μ¡¸Ê¦¤¥´¨¥ ¶μ²ÊÎ¥´´ÒÌ
·¥§Ê²ÓÉ Éμ¢.
‹ˆ’…’“
1. Devin A. B., Prosvirova T. Yu., Peshekhonov V. T., Chepurnaya O. V., Smirnova M. Ye.,
Koltovaya N. A., Troitskaya E. N., Arman I. P. The Start gene CDC28 and the genetic
stability of yeast // Yeast. 1990. V. 6. P. 231Ä243.
2. Koltovaya N. A., Arman I. P., Devin A. B. Mutation of the CDC28 gene and the radiation sensitivity of Saccharomyces cerevisiae // Yeast. 1998. V. 14. P. 133Ä146.
26
3. Koltovaya N. A., Guerasimova A. S., Tchekhouta I. A., Devin A. B. NET1 and HFI1
genes of yeast mediate both chromosome maintenance and mitochondrial rho− mutagenesis // Yeast. 2003. V. 20. P. 955Ä971.
4. Weinert T. A., Hartwell L. H. The RAD9 gene controls the cell cycle response to DNA
damage in Saccharomyces cerevisiae // Science. 1988. V. 241. P. 317Ä322.
5. Chiolo I., Carotenuto W., Mafˇoletti G., Petrini H. J., Foiani M., Liberi G. Srs2
and Sgs1 DNA helicases associate with Mre11 in different subcomplexes following
checkpoint activation and CDK1-mediated Srs2 phosphorylation // Mol. Cell. Biol.
2005. V. 25. P. 5738Ä5751.
6. Michelson R. J., Weinert T. Closing the gaps among a web of DNA repair disorders //
BioEssays. 2000. V. 22. P. 966Ä969.
7. Tercero J. A., Difey J. F. Regulation of DNA replication fork progression through
damaged DNA by the Mec/Rad53 checkpoint // Nature. 2001. V. 412. P. 553Ä557.
8. Painter R. B., Young B. R. Radiosensitivity in ataxia-telangiectasia: a new explanation // Proc. Natl. Acad. Sci. USA. 1980. V. 77. P. 7315Ä7317.
9. Shirahige K., Hori Y., Shiraishi K., Yamashita M., Takahashi K., Obuse C., Tsurimoto
T., Yoshikawa H. Regulation of DNA-replication origins during cell-cycle progression // Nature. 1998. V. 395. P. 618Ä621.
10. Paulovich A. G., Margulies R. U., Garvik B. M., Hartwell L. H. RAD9, RAD17, and
RAD24 are required for S phase regulation in Saccharomyces cerevisiae in response
to DNA damage // Genetics. 1997. V. 145. P. 45Ä62.
11. Lowndes N. F., Murguia J. R. Sensing and responding to DNA damage // Curr. Opin.
Genet. Deversion. 2000. V. 10. P. 17Ä25.
12. Santocanale C., Difey J. F. X. A Mec1- and Rad53-dependent checkpoint controls
late-ˇring origins of DNA replication // Nature. 1998. V. 395. P. 615Ä618.
13. Hartwell L. Defects in a cell cycle checkpoint may be responsible for the genomic
instability of cancer cells // Cell. 1992. V. 71. P. 543Ä546.
14. Zhou B. B., Elledge S. J. The DNA damage response: putting checkpoints in perspective // Nature. 2000. V. 408. P. 433Ä439.
15. Myung K., Datta A., Kolodner R. D. Suppression of spontaneous chromosomal rearrangements by S phase checkpoint functions in Saccharomyces cerevisiae // Cell.
2001. V. 104. P. 397-408.
16. Myung K., Kolodner R. D. Suppression of genome instability by redundant S-phase
checkpoint pathways in Saccharomyces cerevisiae // Proc. Natl. Acad. Sci. USA. 2002.
V. 99. P. 4500Ä4507.
27
17. Šμ²Éμ¢ Ö . ., Š ·¢¨£ ’. „., ‹Õ¡¨³μ¢ Š. ., Œ °μ·μ¢ … .‘., ·³ ´ ˆ. .,
„¥¢¨´ . . ¤¨μÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ ¤·μ¦¦¥° ¸ Ì ·μ³¨Í¥Éμ¢ ¨ £¥´Ò SRM: ÔËË¥±ÉÒ
³ÊÉ Í¨° srm1 ¨ srm5 // ƒ¥´¥É¨± . 1998. ’. 34. ‘. 610Ä624.
18. Šμ²Éμ¢ Ö . ., Š ¤ÒÏ¥¢¸± Ö …. . ƒ¥´ CDC28 ¨ ±μ´É·μ²Ó ±²¥Éμδμ£μ ͨ±² Ê
¤·μ¦¦¥°-¸ Ì ·μ³¨Í¥Éμ¢ // „μ±². . 1997. ’. 357 ‘. 710Ä712.
19. Šμ²Éμ¢ Ö . ., Š ·É Ï¥¢ . ., Š ¤ÒÏ¥¢¸± Ö …. ., —¥ÌÊÉ ˆ. ., ‘¨´¥μ± Ö
ˆ. ‚., ‘³¨·´μ¢ Œ. …., ˜¢ ´¥¢ . ‚., ·³ ´ ˆ. ., „¥¢¨´ . . SRM8 ¨ SRM12 Ä
¤¢ £¥´ RAD9-§ ¢¨¸¨³μ£μ ¶Êɨ, ±μ´É·μ²¨·ÊÕÐ¥£μ · ¤¨μÎÊ¢¸É¢¨É¥²Ó´μ¸ÉÓ Saccharomyces cerevisiae // „μ±². . 1998. ’. 360. ‘. 420Ä422.
20. Šμ²Éμ¢ Ö . ., „¥¢¨´ . . ·μ²¨ ´¥±μÉμ·ÒÌ checkpoint-£¥´μ¢ ¢ μ¶·¥¤¥²¥´¨¨
· ¤¨μÎÊ¢¸É¢¨É¥²Ó´μ¸É¨ ¤·μ¦¦¥° Saccharomyces cerevisiae // „μ±². . 2002.
’. 387. ‘. 1Ä4.
21. Šμ²Éμ¢ Ö . ., ¨±Ê²Êϱ¨´ . ‚., Š ¤ÒÏ¥¢¸± Ö …. ., μШ´ Œ. ., „¥¢¨´
. . ‚§ ¨³μ¤¥°¸É¢¨¥ Î¥±¶μ°´É-£¥´μ¢ RAD9, RAD17, RAD24, RAD53 ¸ £¥´ ³¨
SRM5/CDC28, SRM8/NET1 ¨ SRM12/ HFI1 ¢ μ¶·¥¤¥²¥´¨¨ ÎÊ¢¸É¢¨É¥²Ó´μ¸É¨ ¤·μ¦¦¥° Saccharomyces cerevisiae ± ¤¥°¸É¢¨Õ ¨μ´¨§¨·ÊÕÐ¥° · ¤¨ ͨ¨ // ƒ¥´¥É¨± .
2007. ¢ ¶¥Î.
22. Sherman F., Fink G. R., Hicks J. B. Laboratory course manual for methods in yeast
genetics // Cold Spring Harbor Laboratory. 1986. N. Y.
23. Siede W., Friedberg E. C. Inuence of DNA repair deˇciencies on the UV sensitivity
of yeast cells in different cell cycle stages // Mutation Res. 1990. V. 245. P. 287Ä292.
24. Siede W., Friedberg A. S., Friedberg E. C. RAD9-dependent G1 arrest deˇnes a second
checkpoint for damaged DNA in the cell cycle of Saccharomyces cerevisiae // Proc.
Natl. Acad. Sci. USA. 1993. V. 90. P. 7985Ä7989.
25. Siede W., Friedberg A. S., Dianova I., Friedberg E. C. Characterization of G1 checkpoint control in the yeast Saccharomyces cerevisiae following exposure to DNA
damaging agents // Genetics. 1994. V. 138. P. 271Ä281.
26. Longhese M. P., Frashini R., Plevano P., Lucchini G. Yeast pip3/mec3 mutants fail
to delay entry into S phase and to slow DNA replication in response to DNA damage,
and they deˇne a functional link between Mec3 and DNA primase // Mol. Cell. Biol.
1996. V. 16. P. 3235Ä3244.
27. Allen J. B., Zhou Z., Siede W., Friedberg E. C., Elledge S. J. The SAD1/RAD53
protein kinase controls multiple checkpoints and DNA damage-induced transcription
in yeast // Genes Dev. 1994. V. 8. P. 2401Ä2415.
28. Navas T. A., Sanchez Y., Elledge S. J. RAD9 and DNA polymerase ε form parallel sensory branches for transducing the DNA damage checkpoint signal in Saccharomyces
cerevisiae // Genes Dev. 1996. V. 10. P. 2632Ä2643.
28
29. Weinert T. A., Kiser G. L., Hartwell L. H. Mitotic checkpoint genes in budding yeast
and the dependence of mitosis on DNA replication and repair // Genes Dev. 1994.
V. 8. P. 652Ä665.
30. Johnston L. H., Nasmyth K. A. Saccharomyces cerevisiae cell cycle mutant cdc9 is
defective in DNA ligase // Nature. 1978. V. 274. P. 981Ä893.
31. Weinert T. A., Hartwell L. H. Cell cycle arrest of cdc mutants and speciˇcity of the
RAD9 checkpoint // Genetics. 1993. V. 134. P. 63Ä80.
32. Li X., Cai M. Inactivation of the cyclin-dependent kinase Cdc28 abrogates cell cycle
arrest induced by DNA damage and disassembly of mitotic spindles in Saccharomyces
cerevisiae // Mol. Cell. Biol. 1997. V. 17. P. 2723Ä2734.
33. Ira G., Pellicioli A., Balijja A., Wang X., Fiorani S., Carotenuto W., Liberi G.,
Bressan D., Wan L., Hollingsworth N. M., Haber J. E., Foiani M. DNA end resection,
homologous recombination and DNA damage checkpoint activation require CDK1 //
Nature. 2004. V. 431. P. 1011Ä1017.
34. Mendenhall M. D, Hodge A. E. Regulation of Cdc28 cyclin-dependent protein kinase
activity during the cell cycle of the yeast // Microbiol. Mol. Biol. Rev. 1998. V. 62.
P. 1191Ä1243.
35. Wittenberg C., Reed S. I. Control of the yeast cell cycle is associated with assembly/disassembly of the Cdc28 protein kinase complex // Cell. 1988. V. 54. P. 1061Ä
1072.
36. Mendenhall M. D., Jones C. A., Reed S. I. Dual regulation of the yeast CDC28-p40
protein kinase complex: cell cycle, pheromone, and nutrient limitation effects // Cell.
1987. V. 50. P. 927Ä935.
37. Langan T. A., Gautier J., Lohka M., Hollingsworth R., Moreno S., Nurse P., Maller
J., Sclafani R. A. Mammalian growth-associated H1 histone kinase: a homolog of
cdc2+ /CDC28 protein kinases controlling mitotic entry in yeast and frog cells // Mol.
Cell. Biol. 1989. V. 9. P. 3860Ä3868.
38. Hadwiger J. A., Reed S. I. Invariant phosphorylation of the Saccharomyces cerevisiae
Cdc28 protein kianse // Mol. Cell. Biol. 1988. V. 8. P. 2976Ä2979.
39. Amon A., Surana U., Muroff I., Nasmyth K. Regulation of p34CDC28 tyrosine phosphorylation is not required for entry into mitosis in S. cerevisiae // Nature. 1992.
V. 355. P. 368Ä371.
40. Wittenberg C., Sugimoto K., Reed S. I. G1-speciˇc cyclins of S. cerevisiae: cell cycle
periodicity, regulation by mating pheromone, and association with the p34CDC28
protein kinase // Cell. 1990. V. 62. P. 225Ä37.
41. Cross F. R., Lewin K. Molecular evolution allows bypass of the requirement for
activation loop phosphorylation of the Cdc28 cyclin-dependent kinase // Mol. Cell.
Biol. 1998. V. 18. P. 2823Ä2931.
29
42. Krek W., Nigg E. A. Mutations of p34cdc2 phosphorylation sites induce premature
mitotic events in HeLa cells: evidence for a double block to p34cdc2 kinase activation
in vertebrates // EMBO J. 1991. V. 10. P. 3331Ä3341.
43. Sorger P. K., Murray A. W. S-phase feedback control in budding yeast independent of
tyrosine phosphorylation of p34cdc28 // Nature. 1992. V. 355. P. 365Ä368.
44. Russell P., Moreno S., Reed S. I. Conservation of mitotic controls in ˇssion and
budding yeasts // Cell. 1989. V. 57. P. 295Ä303.
45. Lim H. H., Surana U. Cdc20, a beta-transducin homologue, links RAD9-mediated
G2/M checkpoint control to mitosis in Saccharomyces cerevisiae // Mol. Gen. Genet.
1996. V. 253. P. 138Ä148.
46. Gould K. L., Nurse E. A. Tyrosine phosphorylation of the ˇssion yeast cdc2+ protein
kinase regulates entry into mitosis // Nature. 1989. V. 342. P. 39Ä45.
47. McMillan J. N., Sia R. A. L., Bardes E. S. G., Lew D. J. Phosphorylation-independent
inhibition of Cdc28p by the tyrosine kinase Swe1p in the morphogenesis checkpoint //
Mol. Cell. Biol. 1999. V. 19. P. 5981Ä5990.
48. Alexander M. R., Tyers M., Perret M., Craig B. M., Fang K. S., Gustin M. C. Regulation
of cell cycle progression by Swe1p and Hog1p following hypertonic stress // Mol.
Biol. Cell. 2001. V. 12. P. 53Ä62.
49. Booher R. N., Alfa C. E., Hyams J. S., Beach D. H. The ˇssion yeast cdc2/cdc13/suc1
protein kinase: regulation of catalytic activity and nuclear localization // Cell. 1989.
V. 58. P. 485Ä497.
50. Moreno S., Hayles J., Nurse P. Regulation of p34cdc2 protein kinase during mitosis //
Cell. 1989. V. 58. P. 361Ä372.
51. Hagan I. M., Hyams J. S. The use of cell division cycle mutants to investigate the
control of microtubule distribution in the ˇssion yeast Schizosaecharomyces pombe //
J. Cell Sci. 1988. V. 89. P. 343Ä357.
52. Byer B., Goetsch L. Duplication of spindle plaques and integration of the yeast cell
cycle // Cold Spring Harbor Symp. Quant. Biol. 1974. V. 38. P. 123Ä131.
53. Pringle J. R., Hartwell L. H. The Saccharomyces cerevisiae cell cycle, In: The molecular biology of the yeast Saccharomyces. Life cycle and inheritance. Strathern JN,
Jones EW, Broach JR (Eds.). Cold Spring Harbor Laboratory. Cold Spring Harbor.
NY. 1981. P. 97Ä142.
54. Reed S. I., Wittenberger C. Mitotic role for the Cdc28 protein kinase of Saccharomyces
cerevisiae // Proc. Natl. Acad. Sci. USA. 1990. V. 87. P. 5697Ä5701.
55. Surana U., Robitsch H., Price C., Schuster T., Fitcher A. B., Nasmyth K. The role of
CDC28 and cyclins during mitosis in the budding yeast S. cerevisiae // Cell. 1991.
V. 65. P. 145Ä161.
30
56. Lew D. J., Reed S. I. Morphogenesis in the yeast cell cycle: regulation by Cdc28 and
cyclins // J. Cell Biol. 1993. V. 120. P. 1305Ä1320.
57. Padmashree C. G., Surana U. Cdc28-Clb mitotic kinase negatively regulates bud site
assembly in the budding yeast // J. Cell Sci. 2001. V. 114. P. 207Ä218.
58. Sheu Y. J., Barral Y., Snyder M. Polarized growth controls cell shape and bipolar bud
site selection in Saccharomyces cerevisiae // Mol. Cell. Biol. 2000. V. 20. P. 5235Ä
5247.
59. Ahn S. H., Acurio A., Kron S. J. Regulation of G2/M progression by the STE mitogenactivated protein kinase pathway in budding yeast ˇlamentous growth // Mol. Biol.
Cell. 1999. V. 10. P. 3301Ä3316.
60. Lim H. H., Goh P.-Y., Surana U. Spindle poly body separation in Saccharomyces
cerevisiae requires dephosphorylation of the tyrosine 19 residue of Cdc28 // Mol.
Cell. Biol. 1996. V. 16. P. 6385Ä6397.
61. Š·¥Éμ¢ „. ., •μ²³Ê·μ¤μ¢ •. ’., Šμ²Éμ¢ Ö . . ´ ²¨§ ¸¢Ö§¥° ³¥¦¤Ê ’” ¨
± É ²¨É¨Î¥¸±μ° ¸Ê¡Ñ¥¤¨´¨Í¥° ±¨´ §Ò (´ ɨ¢´μ° ¨ ³ÊÉ ´É´μ°) ¸ ¶μ³μÐÓÕ Œ„³μ¤¥²¨·μ¢ ´¨Ö ±·¨¸É ²²¨Î¥¸±μ° ·¥Ï¥É±¨ ±É¨¢´μ° ±¨´ §Ò CDK2 // †Ê·. ”¨§.
•¨³. 2007. É. 10. ‘. 1Ä6.
62. Stueland C. S., Lew C. S., Cismowski M. J., Reed S. I. Full activation of p34CDC28
histone H1 kinase activity is unable to promote entry into mitosis in checkpointarrested cells of the yeast Saccharomyces cerevisiae // Mol. Cell. Biol. 1993. V. 13.
P. 3744Ä3755.
63. Hadwiger J. A., Wittenberg C., Mendenhall M. D., Reed S. I. The Saccharomyces cerevisiae CKS1 gene, a homolog of the Schizosacccharomyces pombe suc1+ gene, encodes a subunit of the Cdc28 protein kinase complex // Mol. Cell. Biol. 1989. V. 9.
P. 2034Ä2041.
64. Kusubata M., Tokui T., Matsuoka Y., Okumura E., Tachibana K., Hisanaga S., Kishimoto T., Yasuda H., Kamijo M., Ohba Y., Tsujimura K., Yatani R., Inagaki M. p31suc1
suppresses the catalytic function of p34cdc2 kinase for intermediate ˇlament proteins,
in vivo // J. Biol. Chem. 1992. V. 267. P. 20937Ä20942.
65. Tang Y., Reed S. I. The Cdk-associated protein Cks1 functions both in G1 and G2 in
Saccharomyces cerevisiae // Genes Dev. 1993. V. 7. P. 822Ä832.
66. Ubersax J. A., Woodbury E. L., Quang P. N., Paraz M., Blethrow J. D., Shah K., Shokat
K. M., Morgan D. O. Targets of the cyclin-dependent kinase Cdk1 // Nature. 2003.
V. 425. P. 859Ä864.
67. Toh G. W., Lowndes N. F. Role of the Saccharomyces cerevisiae Rad9 protein in
sensing and responding to DNA damage // Biochem. Soc. Trans. 2003. V. 31. P. 242Ä
246.
31
68. Masumoto H., Muramatsu S., Kamimura Y., Araki H. S-Cdk-dependent phosphorylation of Sld2 essential for chromosomal DNA replication in budding yeast // Nature.
2002. V. 415. P. 651Ä655.
69. Lee S. E., Moore J. K., Holmes A., Umezu K., Kolodner R. D., Haber J. E. Ku70,
Mre11/Rad50 and RPA proteins regulate adaptation to G2/M arrest after DNA damage // Cell. 1998. V. 94. P. 399Ä409.
70. Lee S. E., Paques F., Sylvan J., Haber J. E. Role of yeast SIR genes and mating type
in channeling double-strand breaks to homologous and nonhomologous recombination
pathways // Curr .Biol. 1999. V. 9. P. 767Ä770.
71. Sanchez Y., Bachant J., Wang H., Hu F., Liu D., Tetzlaff M., Elledge S. J. Control
of the DNA damage checkpoint by chk1 and rad53 protein kinases through distinct
mechanisms // Science. 1999. V. 286. P. 1166Ä1171.
72. Liberi G., Chiolo I., Pellicioli A., Lopes M., Plevani P., Muzi-Falconi M., Foiani
M. Srs2 DNA helicase is involved in checkpoint response and its regulation requires
a functional Mec1-dependent pathway and Cdk1 activity // EMBO J. 2000. V. 19.
P. 5027Ä5038.
73. Horiuchi J., Silverman N., Pina B., Marcus G. A., Guarente L. ADA1, a novel component of the ADA/GCN5 complex, has broader effects than GCN5, ADA2, or ADA3 //
Mol. Cell. Biol. 1997. V. 17. P. 3220Ä3228.
74. Imoberdorf R. M., Topalidou I., Strubin M. A role for Gcn5-mediated global histone
acetylation in transcriptional regulation // Mol. Cell. Biol. 2006. V. 26. P. 1610Ä1616.
75. Megee P. C., Morgan B. A., Mittman B. A., Smith M. M. Genetic analysis of histone
H4: essential role of lysines subject to reversible acetylation // Science. 1990. V. 247.
P. 841Ä845.
76. Megee P. C., Morgan B. A., Smith M. M. Histone H4 and the maintenance of genome
integrity // Genes Dev. 1995. V. 9. P. 1716Ä1727.
77. Clarke A. S., Lowell J. E., Jacobson S. J., Pillus L. Esa1p is an essensial histone
acetyltransferase required for cell cycle progression // Mol. Cell. Biol. 1999. V. 19.
P. 2515Ä2526.
78. Scott K. L., Plon S. E. Loss of Sin3/Rpd3 histone deacetylase restores the DNA damage
response in checkpoint-deˇcient strains of Saccharomyces cerevisiae // Mol. Cell. Biol.
2003. V. 23. P. 4522Ä4531.
79. Aparicio J. G., Viggiani C. J., Gibson D. G., Aparicio O. M. The Rpd3-Sin3 histone
deacetylase regulates replication timing and enables intra-S origin control in Saccharomyces cerevisiae // Mol. Cell. Biol. 2004. V. 24. P. 4769Ä4780.
80. Sakaguchi K., Herrera J. E., Saito S., Miki T., Bustin M., Vassilev A., Anderson
C. W., Appella E. DNA damage activates p53 through a phosphorylation-acetylation
cascade // Genes Dev. 1998. V. 12. P. 2831Ä2841.
32
81. Liu L., Scolnick D. M., Trievel R. C., Zhang H. B., Marmorstein R., Halazonetis T. D.,
Berger S. L. p53 sites acetylated in vitro by PCAF and p300 are acetylated in vivo in
response to DNA damage // Mol. Cell. Biol. 1999. V. 19. P. 1202Ä1209.
82. Barlev N. A., Liu L., Chehab N. H., Mansˇeld K., Harris K. G., Halazonetis T. D.,
Berger S. L. Acetylation of p53 activates transcription through recruitment of coactivators/histone acetytransferases // Mol. Cell. 2001. V. 8. P. 1243Ä1254.
83. Luo J., Nikolaev A. Y., Imai S., Chen D., Su F., Shiloh A., Guarente L., Gu W.
Negative control of p53 by Sir2α promotes cell survival under stress // Cell. 2001.
V. 107. P. 137Ä148.
84. Vaziri H., Dessain S. K., Ng Eaton E., Imai S. I., Frye R. A., Pandita T. K., Guarente
L., Weinberg R. A. hSIR2 (SIRT1) functions as an NAD-dependent p53 deacetylase //
Cell. 2001. V. 107. P. 149Ä159.
85. Wang T., Kobayashi T., Takimoto R., Denes A. E., Snyder E. L., el-Deiry W. S., Brachmann R. K. hADA3 is required for p53 activity // EMBO J. 2001. V. 20. P. 6404Ä6413.
86. Gu W., Roeder R. Activation of p53 sequence-speciˇc DNA binding by acetylation of
the p53 C-terminal domain // Cell. 1997. V. 90. P. 595Ä606.
87. Liu D., Ishima R., Tong K. I., Bagby S., Kokubo T., Muhandiram D. R., Kay L. E.,
Nakataki Y., Ikira M. Solution of a TBP-TAFII 230 complex: protein mimicry of
the minor groove surface of the TATA box unwound by TBP // Cell. 1998. V. 94.
P. 573Ä583.
88. Lo W. S., Duggan L., Emre N. C., Belotserkovskya R., Lane W. S., Shiekhattar R.,
Berger S. L. Snf1 Å a histone kinase that works in concert with the histone acetyltransferase Gcn5 to regulate transcription // Science. 2001. V. 293. P. 1142Ä1146.
89. Clements A., Poux A. N., Lo W. S., Pillus L., Berger S. L., Marmorstein R. Structural
basis for histone and phosphohistone binding by the GCN5 histone acetyltransferase //
Mol. Cell. 2003. V. 12. P. 461Ä473.
90. John S., Howe L., Tafrov S. T., Grant P. A., Sternglanz R., Workman J. L. The something about silencing protein, Sas3, is the catalytic subunit of NuA3, a yTAF(II)30containing HAT complex that interacts with the Spt16 subunit of the yeast CP
(Cdc68/Pob3)-FACT complex // Genes Dev. 2000. V. 14. P. 1196Ä208.
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