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’…’“
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