Cell growth-regulated expression of mammalian MCM5 and MCM6 genes mediated by the transcription factor E2F

Initiation of DNA replication requires the function of MCM gene products, which participate in ensuring that DNA replication occurs only once in the cell cycle. Expression of all mammalian genes of the MCM family is induced by growth stimulation, unlike yeast, and the mRNA levels peak at G1/S boundary. In this study, we examined the transcriptional activities of isolated human MCM gene promoters. Human MCM5 and MCM6 promoters with mutation in the E2F sites failed in promoter regulation following serum stimulation and exogenous E2F expression. In addition, we identified a novel E2F-like sequence in human MCM6 promoter which cooperates with the authentic E2F sites in E2F-dependent regulation. Forced expression of E2F1 could induce expression of all members of the endogenous MCM genes in rat embryonal fibroblast REF52 cells. Our results demonstrated that the growth-regulated expression of mammalian MCM5 and MCM6 genes, and presumably other MCM members, is primarily regulated by E2F through binding to multiple E2F sites in the promoters.

[1]  M. Fujita,et al.  Inhibition of S-phase entry of human fibroblasts by an antisense oligomer against hCDC47. , 1996, Biochemical and biophysical research communications.

[2]  P. Rigby,et al.  The embryonal carcinoma stem cell Ela-like activity involves a differentiation-regulated transcription factor. , 1990, Nucleic acids research.

[3]  W. Kaelin,et al.  Deregulated transcription factor E2F-1 expression leads to S-phase entry and p53-mediated apoptosis. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[4]  K. Arai,et al.  Human and Xenopus cDNAs encoding budding yeast Cdc7‐related kinases: in vitro phosphorylation of MCM subunits by a putative human homologue of Cdc7 , 1997, The EMBO journal.

[5]  Y. Endo,et al.  HsMCM6: a new member of the human MCM/P1 family encodes a protein homologous to fission yeast Mis5 , 1997, Genes to cells : devoted to molecular & cellular mechanisms.

[6]  J. Nevins,et al.  Expression of the HsOrc1 gene, a human ORC1 homolog, is regulated by cell proliferation via the E2F transcription factor , 1996, Molecular and cellular biology.

[7]  H. Nojima,et al.  Licensing of DNA replication by a multi‐protein complex of MCM/P1 proteins in Xenopus eggs , 1997, The EMBO journal.

[8]  E. Lam,et al.  An E2F‐binding site mediates cell‐cycle regulated repression of mouse B‐myb transcription. , 1993, The EMBO journal.

[9]  Y. Endo,et al.  Expression, nuclear localization and interactions of human MCM/P1 proteins. , 1997, Biochemical and biophysical research communications.

[10]  J. Nevins,et al.  The accumulation of an E2F-p130 transcriptional repressor distinguishes a G0 cell state from a G1 cell state , 1996, Molecular and cellular biology.

[11]  J. Nevins,et al.  The adenovirus-inducible factor E2F stimulates transcription after specific DNA binding , 1989, Molecular and cellular biology.

[12]  L. Drury,et al.  Cdc6p-dependent loading of Mcm proteins onto pre-replicative chromatin in budding yeast. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[13]  J. Nevins,et al.  Expression of the E2F1 transcription factor overcomes type beta transforming growth factor-mediated growth suppression. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[14]  W. Lee,et al.  Deregulated expression of E2F-1 induces S-phase entry and leads to apoptosis , 1994, Molecular and cellular biology.

[15]  J. Nevins,et al.  E2F: a link between the Rb tumor suppressor protein and viral oncoproteins. , 1992, Science.

[16]  M. Fujita,et al.  Cloning of a cDNA encoding a human homologue of CDC47, a member of the MCM family. , 1996, Biochimica et biophysica acta.

[17]  D. Botstein,et al.  [9] Construction and use of gene fusions to lacZ (β-galactosidase) that are expressed in yeast , 1983 .

[18]  J. Nevins,et al.  A unique role for the Rb protein in controlling E2F accumulation during cell growth and differentiation. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[19]  K. Nasmyth,et al.  Loading of an Mcm Protein onto DNA Replication Origins Is Regulated by Cdc6p and CDKs , 1997, Cell.

[20]  Kiyoshi Ohtani,et al.  Regulation of cell growth-dependent expression of mammalian CDC6 gene by the cell cycle transcription factor E2F , 1998, Oncogene.

[21]  K. Sugimoto,et al.  DNA polymerase alpha associated protein P1, a murine homolog of yeast MCM3, changes its intranuclear distribution during the DNA synthetic period. , 1994, The EMBO journal.

[22]  J. Nevins,et al.  Expression of transcription factor E2F1 induces quiescent cells to enter S phase , 1993, Nature.

[23]  J. Nevins,et al.  Distinct roles for E2F proteins in cell growth control and apoptosis. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[24]  James P. J. Chong,et al.  Purification of an MCM-containing complex as a component of the DNA replication licensing system , 1995, Nature.

[25]  R. Laskey,et al.  MCM3 complex required for cell cycle regulation of DNA replication in vertebrate cells , 1995, Nature.

[26]  J. Nevins,et al.  Autoregulatory control of E2F1 expression in response to positive and negative regulators of cell cycle progression. , 1994, Genes & development.

[27]  H. Nojima,et al.  Identification of the yeast MCM3-related protein as a component of xenopus DNA replication licensing factor , 1995, Cell.

[28]  J. Nevins,et al.  Cdc6 is regulated by E2F and is essential for DNA replication in mammalian cells. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[29]  J. Blow,et al.  A role for the nuclear envelope in controlling DNA replication within the cell cycle , 1988, Nature.

[30]  N. L. Thangue,et al.  DRTF1/E2F: an expanding family of heterodimeric transcription factors implicated in cell-cycle control. , 1994 .

[31]  J. Blow,et al.  The RLF‐M component of the replication licensing system forms complexes containing all six MCM/P1 polypeptides , 1997, The EMBO journal.