The RLF-M component of the replication licensing system forms complexes containing all six MCM/P1 polypeptides

Thömmes, Pia, Kubota, Yumiko, Takisawa, Haruhiko and Blow, J. Julian (1997) The RLF-M component of the replication licensing system forms complexes containing all six MCM/P1 polypeptides. EMBO Journal, 16. pp. 3312-3319. ISSN 0261-4189

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Abstract

Replication licensing factor (RLF) is involved in preventing re-replication of chromosomal DNA in a single cell cycle, and previously has been separated into two components termed RLF-M and RLF-B. Here we show that Xenopus RLF-M consists of all six members of the MCM/P1 protein family, XMcm2-XMcm7. The six MCM/P1 polypeptides co-eluted on glycerol gradients and gel filtration as complexes with a mol. wt of approximately 400 kDa. In crude Xenopus extract, all six MCM/P1 polypeptides co-precipitated with anti-XMcm3 antibody, although only XMcm5 quantitatively co-precipitated from purified RLF-M. Further fractionation separated RLF-M into two sub-components, one consisting of XMcms 3 and 5, the other consisting of XMcms 2, 4, 6 and 7. Neither of the sub-components provided RLF-M activity. Finally, we show that all six MCM/P1 proteins bind synchronously to chromatin before the onset of S-phase and are displaced as S-phase proceeds. These results strongly suggest that complexes containing all six MCM/P1 proteins are necessary for replication licensing.

Item Type:	Article
Faculty \ School:
Depositing User:	LivePure Connector
Date Deposited:	11 Jun 2024 09:30
Last Modified:	11 Jun 2024 09:30
URI:	https://ueaeprints.uea.ac.uk/id/eprint/95540
DOI:	10.1093/emboj/16.11.3312

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