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MCP1 与哺乳动物细胞中复制机制成分的相互作用。

Interactions of MCP1 with components of the replication machinery in mammalian cells.

机构信息

Departamento de Ciências Biológicas, Laboratório de Bioquímica, Faculdade de Farmácia da Universidade do Porto, Portugal.

出版信息

Int J Biol Sci. 2011 Feb 17;7(2):193-208. doi: 10.7150/ijbs.7.193.

DOI:10.7150/ijbs.7.193
PMID:21383955
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3048848/
Abstract

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.

摘要

真核生物的 DNA 复制从复制起点预复制复合物 (pre-RC) 的组装开始。我们之前已经证明，中期染色体蛋白 1 (MCP1) 参与了 DNA 复制的早期事件。在这里，我们表明 MCP1 与建立预复制复合物所需的蛋白质相关。相互免疫沉淀分析表明，MCP1 与 Cdc6、ORC2、ORC4、MCM2、MCM3 和 MCM7 与 Cdc45 和 PCNA 相互作用。免疫荧光研究表明 MCP1 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，MCP1 优先结合人细胞中的复制起始位点。有趣的是，尽管预 RC 的成员已知与异染色质的某些特征相互作用，但我们的共免疫沉淀和免疫荧光分析表明 MCP1 不与包括 HP1β 和 MetH3K9 在内的异染色质蛋白相互作用，也不共定位。这些观察结果表明，MCP1 与复制因子相关联，这些因子是启动 DNA 复制所必需的，并且与在 S 期早期复制的基因座中的起始位点结合。此外，免疫测定法揭示了 MCP1 形式与组蛋白 H1 变体的关联，质谱分析证实 MCP1 肽与 H1.2 和 H1.5 亚型共享共同序列。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/8ea4c756f8c8/ijbsv07p0193g10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/4c247ec9e5e6/ijbsv07p0193g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/1a28c2b34891/ijbsv07p0193g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/ead5582184ce/ijbsv07p0193g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/d8ba8a5bf013/ijbsv07p0193g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/27ef3e9ff488/ijbsv07p0193g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/90146443c844/ijbsv07p0193g06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/fd3511667227/ijbsv07p0193g07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/94cb89e4e944/ijbsv07p0193g08.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/acb6cd6d1a6d/ijbsv07p0193g09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/8ea4c756f8c8/ijbsv07p0193g10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/4c247ec9e5e6/ijbsv07p0193g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/1a28c2b34891/ijbsv07p0193g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/ead5582184ce/ijbsv07p0193g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/d8ba8a5bf013/ijbsv07p0193g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/27ef3e9ff488/ijbsv07p0193g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/90146443c844/ijbsv07p0193g06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/fd3511667227/ijbsv07p0193g07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/94cb89e4e944/ijbsv07p0193g08.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/acb6cd6d1a6d/ijbsv07p0193g09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e486/3048848/8ea4c756f8c8/ijbsv07p0193g10.jpg

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本文引用的文献

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