CDK-INTERACTING PROTEIN-1 DIRECTLY BINDS WITH PROLIFERATING CELL NUCLEAR ANTIGEN AND INHIBITS DNA-REPLICATION CATALYZED BY THE DNA-POLYMERASE-DELTA HOLOENZYME
CDK-INTERACTING PROTEIN-1 DIRECTLY BINDS WITH PROLIFERATING CELL NUCLEAR ANTIGEN AND INHIBITS DNA-REPLICATION CATALYZED BY THE DNA-POLYMERASE-DELTA HOLOENZYME
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DOI：
10.1073/pnas.91.18.8655
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发表时间：
1994-08-30
影响因子：
11.1
通讯作者：
HURWITZ, J
中科院分区：
文献类型：
--
作者：
FLORESROZAS, H;KELMAN, Z;HURWITZ, J
Cdk相互作用蛋白1（Cip 1）是一种p53调节的21-kDa蛋白，其抑制细胞周期蛋白依赖性激酶（CDK）家族的几个成员。最初在含有CDK 4、细胞周期蛋白D和增殖细胞核抗原（PCNA）的复合物中观察到。PCNA与激活因子1一起作为真核生物DNA聚合酶（pol）δ的持续合成因子，这三种蛋白质构成pol δ全酶。在这份报告中，我们表明，Cip 1也可以直接抑制DNA合成在体外结合PCNA。Cip 1有效抑制依赖于pol α、激活因子1、PCNA和pol δ的Sinian病毒40复制，并且这种抑制可以被额外的PCNA克服。猿猴病毒40 DNA复制，仅由高水平的pol α-引发酶复合物催化，不受Cip 1的影响。使用表面等离子体共振技术，检测到PCNA和Cip 1的直接物理相互作用。我们已经观察到，Cip 1有效地抑制长（7.2 kb），但不短（10 nt）模板的合成，这表明它与PCNA的关联可能会损害在DNA链延伸过程中的pol δ的进行性运动，而不是阻止组装的pol δ全酶。Cip 1-PCNA相互作用的影响与调节DNA合成，细胞周期检查点控制，和DNA修复进行了讨论。
Cdk-interacting protein 1 (Cip1) is a p53-regulated 21-kDa protein that inhibits several members of the cyclin-dependent kinase (CDK) family. It was initially observed in complexes containing CDK4, cyclin D, and proliferating cell nuclear antigen (PCNA). PCNA, in conjunction with activator 1, acts as a processivity factor for eukaryotic DNA polymerase (pol) delta, and these three proteins constitute the pol delta holoenzyme. In this report, we demonstrate that Cip1 can also directly inhibit DNA synthesis in vitro by binding to PCNA. Cip1 efficiently inhibits sinian virus 40 replication dependent upon pol alpha, activator 1, PCNA, and pol delta, and this inhibition can be overcome by additional PCNA. Simian virus 40 DNA replication, catalyzed solely by high levels of pol alpha-primase complex, is unaffected by Cip1. Using the surface plasmon resonance technique, a direct physical interaction of PCNA and Cip1 was detected. We have observed that Cip1 efficiently inhibits synthesis of long (7.2 kb) but not short (10 nt) templates, suggesting that its association with PCNA is likely to impair the processive movement of pol delta during DNA chain elongation, as opposed to blocking assembly of the pol delta holoenzyme. The implications of the Cip1-PCNA interaction with respect to regulation of DNA synthesis, cell cycle checkpoint control, and DNA repair are discussed.