| Budget Amount *help |
¥3,980,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥480,000)
Fiscal Year 2007: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Research Abstract |
Coordinated execution of DNA replication, checkpoint activation and post-replicative chromatid cohesion is intimately related to the replication fork machinery. In this study, we focused on the human homologs of replication complex components, AND-1, Tim, and Tipin and identified their functions in S phase as below.
1. Human Tim and Tipin form complex during cell cycle. Coimmunoprecipitation experiments reveals that Tim also interacts with Mcm2 and AND-1 interacts with cohesin Smc1, Smc3 Rad21 and Mcm7.
2. AND-1, Tim and Tipin localize adjacent to replication foci and are required for efficient DNA replication.
3. Depletion of AND-1 causes DNA damage and cell death.
4. AND-1, Tim and Tipin are required for efficient replication checkpoint and promote UV-resistant DNA replication. Tim and Tipin are also required for chromatin loading of Claspin in replication stress.
5. AND-1 is phosphorylated by replication arrest which depends on ATM/ATR and Cdc7 kinase.
6. AND-1 is required for efficient recombinational repair.
Based on these data, we propose that AND- 1, Tim and Tipin forms complexes with cohesin and replication factors and coordinates multiple cellular events in S phase, such as DNA replication, checkpoint activation, sister chromatid cohesion and repair, thus playing a pivotal role in maintenance of genome integrity.
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