Interactions of MCP1 with components of the replication machinery in mammalian cells.
Interactions of MCP1 with components of the replication machinery in mammalian cells.
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DOI：
10.7150/ijbs.7.193
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发表时间：
2011-02-17
影响因子：
9.2
通讯作者：
Aladjem MI
中科院分区：
文献类型：
--
作者：
Bronze-da-Rocha E;Lin CM;Shimura T;Aladjem MI
真核DNA复制开始于复制起点处的复制前复合物（pre-RC）的组装。我们以前已经证明，中期染色体蛋白1（MCP 1）参与DNA复制的早期事件。在这里，我们表明，MCP 1与建立复制前复合物所需的蛋白质相关联。免疫共沉淀分析显示MCP 1与Cdc 6、ORC 2、ORC 4、MCM 2、MCM 3和MCM 7相互作用，与Cdc 45和PCNA相互作用。免疫荧光研究表明MCP 1与其中一些蛋白质共定位。此外，利用染色质免疫沉淀（ChIP）的生化研究表明，MCP 1优先结合人类细胞中的复制起始位点。有趣的是，尽管已知前RC的成员与异染色质的一些标志物相互作用，但我们的免疫共沉淀和免疫荧光分析表明MCP 1不与异染色质蛋白（包括HP 1 β和MetH 3 K9）相互作用，也不与异染色质蛋白共定位。这些观察结果表明，MCP 1与DNA复制启动所需的复制因子相关，并结合到S期早期复制的基因座中的起始位点。此外，免疫学测定揭示了MCP 1形式与组蛋白H1变体的关联，质谱分析证实MCP 1肽与H1.2和H1.5亚型共享共同序列。
Eukaryotic DNA replication starts with the assembly of a pre-replication complex (pre-RC) at replication origins. We have previously demonstrated that Metaphase Chromosome Protein 1 (MCP1) is involved in the early events of DNA replication. Here we show that MCP1 associates with proteins that are required for the establishment of the pre-replication complex. Reciprocal immunoprecipitation analysis showed that MCP1 interacted with Cdc6, ORC2, ORC4, MCM2, MCM3 and MCM7, with Cdc45 and PCNA. Immunofluorescence studies demonstrated the co-localization of MCP1 with some of those proteins. Moreover, biochemical studies utilizing chromatin-immunoprecipitation (ChIP) revealed that MCP1 preferentially binds replication initiation sites in human cells. Interestingly, although members of the pre-RC are known to interact with some hallmarks of heterochromatin, our co-immunoprecipitation and immunofluorescence analyses showed that MCP1 did not interact and did not co-localize with heterochromatic proteins including HP1β and MetH3K9. These observations suggest that MCP1 is associated with replication factors required for the initiation of DNA replication and binds to the initiation sites in loci that replicate early in S-phase. In addition, immunological assays revealed the association of MCP1 forms with histone H1 variants and mass spectrometry analysis confirmed that MCP1 peptides share common sequences with H1.2 and H1.5 subtypes.
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影响因子：
9.2
作者：
Lin, CM;Fu, HQ;Aladjem, MI
通讯作者：
Aladjem, MI
DOI：
10.1073/pnas.85.21.8081
发表时间：
1988-11-01
影响因子：
11.1
作者：
EPNER, E;FORRESTER, WC;GROUDINE, M
通讯作者：
GROUDINE, M
影响因子：
2.6
作者：
Bronze-da-Rocha, E;Catita, JA;Sunkel, CE
通讯作者：
Sunkel, CE
影响因子：
56.9
作者：
Festenstein, R;Pagakis, SN;Kioussis, D
通讯作者：
Kioussis, D
DOI：
10.1083/jcb.111.4.1519
发表时间：
1990-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Bernat RL;Borisy GG;Rothfield NF;Earnshaw WC
通讯作者：
Earnshaw WC