Site-specific phosphorylation of MCM4 during the cell cycle in mammalian cells

The FEBS Journal
Yuki Komamura-KohnoYukio Ishimi

Abstract

MCM4, a subunit of a putative replicative helicase, is phosphorylated during the cell cycle, at least in part by cyclin-dependent kinases (CDK), which play a central role in the regulation of DNA replication. However, detailed characterization of the phosphorylation of MCM4 remains to be performed. We examined the phosphorylation of human MCM4 at Ser3, Thr7, Thr19, Ser32, Ser54, Ser88 and Thr110 using anti-phosphoMCM4 sera. Western blot analysis of HeLa cells indicated that phosphorylation of MCM4 at these seven sites can be classified into two groups: (a) phosphorylation that is greatly enhanced in the G2 and M phases (Thr7, Thr19, Ser32, Ser54, Ser88 and Thr110), and (b) phosphorylation that is firmly detected during interphase (Ser3). We present data indicating that phosphorylation at Thr7, Thr19, Ser32, Ser88 and Thr110 in the M phase requires CDK1, using a temperature-sensitive mutant of mouse CDK1, and phosphorylation at sites 3 and 32 during interphase requires CDK2, using a dominant-negative mutant of human CDK2. Based on these results and those from in vitro phosphorylation of MCM4 with CDK2/cyclin A, we discuss the kinases responsible for MCM4 phosphorylation. Phosphorylated MCM4 detected using anti-phospho sera exhib...Continue Reading

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Citations

Jan 20, 2009·The Journal of Biological Chemistry·Min DengXin Ye
Dec 1, 2009·Microbiology and Molecular Biology Reviews : MMBR·Matthew L Bochman, Anthony Schwacha
Dec 13, 2006·Microbiology and Molecular Biology Reviews : MMBR·Elizabeth R Barry, Stephen D Bell
Jul 7, 2007·BMC Bioinformatics·Michelle S Scott, Geoffrey J Barton
Oct 14, 2008·Genome Biology·Yong ChiBruce E Clurman
Jul 19, 2013·Journal of Biochemistry·Mariko Moritani, Yukio Ishimi
Nov 6, 2007·Proceedings of the National Academy of Sciences of the United States of America·Alan M MosesRichard Durbin
Jan 12, 2011·The Journal of Cell Biology·Marjorie A KuipersDavid M Gilbert
Jan 22, 2013·Communicative & Integrative Biology·Abulaish Ansari, Renu Tuteja
Sep 6, 2018·Journal of Biochemistry·Xuan Wang, Yukio Ishimi
Oct 30, 2018·Genes & Genetic Systems·Yukio Ishimi
Feb 14, 2020·Molecular & Cellular Proteomics : MCP·Patrick HerrMikael Altun
Jun 8, 2017·Scientific Reports·Dianpeng ZhengJinlong Li
Aug 1, 2018·Cell & Bioscience·Liangru Fei, Hongtao Xu
Jan 17, 2021·Molecular & Cellular Proteomics : MCP·Patrick HerrMikael Altun

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