Association of Rad 9 with Double-Strand Breaks through a Mec 1-Dependent Mechanism
Association of Rad 9 with Double-Strand Breaks through a Mec 1-Dependent Mechanism
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Rad 9 与双链的结合突破了 Mec 1 依赖性机制
DOI：
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
K. Sugimoto
中科院分区：
文献类型：
--
作者：
T. Naiki;Tatsushi Wakayama;D. Nakada;Kunihiro Matsumoto;K. Sugimoto
Rad 9是芽殖酵母中DNA损伤检查点途径激活所必需的。Rad 9在DNA损伤后以Mec 1和Tel 1依赖的方式磷酸化，随后与Rad 53相互作用。Rad 9与Rad 53的相互作用被认为会触发Rad 53的激活和磷酸化。在这里，我们表明，Mec 1控制Rad 9积累在双链断裂（DSB）。Rad 9在DSB诱导后被磷酸化并与DSB相关。然而，它的磷酸化和与DSB的关联在携带mec 1或激酶阴性mec 1突变的细胞中显著降低。Mec 1在体外磷酸化Rad 9的S/TQ基序，与体内DNA损伤后磷酸化的基序相同。此外，Rad 9 S/TQ基序的多个突变导致其与DSB的结合缺陷。在携带弱mec 1等位基因（mec 1 -81）的细胞中，Rad 9的磷酸化部分缺陷，而在mec 1 -81突变体中，它与DSB的结合有效，正如在野生型细胞中发现的那样。然而，Rad 9-Rad 53的相互作用DSB诱导后显着降低mec 1 -81突变体，因为它是在mec 1突变体。RAD 53缺失突变不影响Rad 9与DSB的关联。我们的研究结果表明，Mec 1促进协会的Rad 9与DNA损伤的网站，从而导致完全磷酸化的Rad 9和Rad 53的相互作用。
Rad9 is required for the activation of DNA damage checkpoint pathways in budding yeast. Rad9 is phosphorylated after DNA damage in a Mec1and Tel1-dependent manner and subsequently interacts with Rad53. This Rad9-Rad53 interaction has been suggested to trigger the activation and phosphorylation of Rad53. Here we show that Mec1 controls the Rad9 accumulation at double-strand breaks (DSBs). Rad9 was phosphorylated after DSB induction and associated with DSBs. However, its phosphorylation and association with DSBs were significantly decreased in cells carrying a mec1 or kinase-negative mec1 mutation. Mec1 phosphorylated the S/TQ motifs of Rad9 in vitro, the same motifs that are phosphorylated after DNA damage in vivo. In addition, multiple mutations in the Rad9 S/TQ motifs resulted in its defective association with DSBs. Phosphorylation of Rad9 was partially defective in cells carrying a weak mec1 allele (mec1-81), whereas its association with DSBs occurred efficiently in the mec1-81 mutants, as found in wild-type cells. However, the Rad9-Rad53 interaction after DSB induction was significantly decreased in mec1-81 mutants, as it was in mec1 mutants. Deletion mutation in RAD53 did not affect the association of Rad9 with DSBs. Our results suggest that Mec1 promotes association of Rad9 with sites of DNA damage, thereby leading to full phosphorylation of Rad9 and its interaction with Rad53.
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DOI：
10.1073/pnas.250475697
发表时间：
2000-12-05
影响因子：
11.1
作者：
Mallory, JC;Petes, TD
通讯作者：
Petes, TD
影响因子：
10.5
作者：
Justine A Melo;J. Cohen;D. Toczyski
通讯作者：
Justine A Melo;J. Cohen;D. Toczyski
影响因子：
56.9
作者：
Sanchez, Y;Bachant, J;Elledge, SJ
通讯作者：
Elledge, SJ
影响因子：
16
作者：
Schwartz, MF;Duong, JK;Stern, DF
通讯作者：
Stern, DF
影响因子：
11.4
作者：
WHITE, CI;HABER, JE
通讯作者：
HABER, JE