Mcm1 Binds Replication Origins*

Mcm1 is an essential protein required for the efficient replication of minichromosomes and the transcriptional regulation of early cell cycle genes in Saccharomyces cerevisiae. In this study, we report that Mcm1 is an abundant protein that associates globally with chromatin in a punctate pattern. We show that Mcm1 is localized at replication origins and plays an important role in the initiation of DNA synthesis at a chromosomal replication origin in vivo. Using purified Mcm1 protein, we show that Mcm1 binds cooperatively to multiple sites at autonomously replicating sequences. These results suggest that, in addition to its role as a transcription factor for the expression of replication genes, Mcm1 may influence the local structure of replication origins by direct binding.

[1]  C. Newlon,et al.  Mcm1 regulates donor preference controlled by the recombination enhancer in Saccharomyces mating-type switching. , 1998, Genes & development.

[2]  B Sugden,et al.  DNA looping between the origin of replication of Epstein-Barr virus and its enhancer site: stabilization of an origin complex with Epstein-Barr nuclear antigen 1. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[3]  A. Feinberg,et al.  A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. , 1983, Analytical biochemistry.

[4]  John J. Wyrick,et al.  Genome-Wide Distribution of ORC and MCM Proteins in S. cerevisiae: High-Resolution Mapping of Replication Origins , 2001, Science.

[5]  B. Tye,et al.  Nuclear accumulation of Saccharomyces cerevisiae Mcm3 is dependent on its nuclear localization sequence , 1997, Genes to cells : devoted to molecular & cellular mechanisms.

[6]  M. Grunstein,et al.  Spreading of transcriptional represser SIR3 from telomeric heterochromatin , 1996, Nature.

[7]  M. Lei,et al.  Initiating DNA synthesis: from recruiting to activating the MCM complex. , 2001, Journal of cell science.

[8]  David M. Gilbert,et al.  Making Sense of Eukaryotic DNA Replication Origins , 2001, Science.

[9]  B. Tye,et al.  Functional domains of the yeast transcription/replication factor MCM1. , 1991, Genes & development.

[10]  R. W. Davis,et al.  Isolation and characterisation of a yeast chromosomal replicator , 1979, Nature.

[11]  G. Costanzo,et al.  ABFI contributes to the chromatin organization of Saccharomyces cerevisiae ARS1 B-domain. , 1994, Biochimica et biophysica acta.

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

[13]  Anindya Dutta,et al.  Replication from oriP of Epstein-Barr Virus Requires Human ORC and Is Inhibited by Geminin , 2001, Cell.

[14]  B. Tye,et al.  Mutants of S. cerevisiae defective in the maintenance of minichromosomes. , 1984, Genetics.

[15]  D. K. Bishop.  RecA homologs Dmc1 and Rad51 interact to form multiple nuclear complexes prior to meiotic chromosome synapsis , 1994, Cell.

[16]  H. Zhong,et al.  DNA-binding specificity of Mcm1: operator mutations that alter DNA-bending and transcriptional activities by a MADS box protein , 1997, Molecular and cellular biology.

[17]  S. Francesconi,et al.  The OBF1 protein and its DNA-binding site are important for the function of an autonomously replicating sequence in Saccharomyces cerevisiae , 1989, Molecular and cellular biology.

[18]  J. Broach,et al.  Localization and sequence analysis of yeast origins of DNA replication. , 1983, Cold Spring Harbor symposia on quantitative biology.

[19]  B. Stillman,et al.  A yeast chromosomal origin of DNA replication defined by multiple functional elements. , 1992, Science.

[20]  L. Breeden,et al.  A novel Mcm1-dependent element in the SWI4, CLN3, CDC6, and CDC47 promoters activates M/G1-specific transcription. , 1997, Genes & development.

[21]  R. Simpson.  Nucleosome positioning can affect the function of a cis-acting DMA elementin vivo , 1990, Nature.

[22]  A. Shevchenko,et al.  Forkhead transcription factors, Fkh1p and Fkh2p, collaborate with Mcm1p to control transcription required for M-phase , 2000, Current Biology.

[23]  R. Elble,et al.  A protein involved in minichromosome maintenance in yeast binds a transcriptional enhancer conserved in eukaryotes. , 1989, Genes & development.

[24]  L. Frappier,et al.  Epstein-Barr nuclear antigen 1 mediates a DNA loop within the latent replication origin of Epstein-Barr virus. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[25]  R. Treisman.  Inside the MADS box , 1995, Nature.

[26]  L. Breeden,et al.  Characterization of the ECB Binding Complex Responsible for the M/G1-Specific Transcription of CLN3 and SWI4 , 2002, Molecular and Cellular Biology.

[27]  Michael Ruogu Zhang,et al.  Comprehensive identification of cell cycle-regulated genes of the yeast Saccharomyces cerevisiae by microarray hybridization. , 1998, Molecular biology of the cell.

[28]  Nicola J. Rinaldi,et al.  Serial Regulation of Transcriptional Regulators in the Yeast Cell Cycle , 2001, Cell.

[29]  M. Grunstein,et al.  Stable nucleosome positioning and complete repression by the yeast alpha 2 repressor are disrupted by amino-terminal mutations in histone H4. , 1992, Genes & development.

[30]  B. Tye,et al.  A family of Saccharomyces cerevisiae repetitive autonomously replicating sequences that have very similar genomic environments. , 1983, Journal of molecular biology.

[31]  M. Lei,et al.  Physical interactions among Mcm proteins and effects of Mcm dosage on DNA replication in Saccharomyces cerevisiae , 1996, Molecular and cellular biology.

[32]  B. Stillman,et al.  Functional conservation of multiple elements in yeast chromosomal replicators , 1994, Molecular and cellular biology.

[33]  C. Newlon,et al.  Two Compound Replication Origins in Saccharomyces cerevisiae Contain Redundant Origin Recognition Complex Binding Sites , 2001, Molecular and Cellular Biology.

[34]  S. Francesconi,et al.  A DNA replication enhancer in Saccharomyces cerevisiae. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[35]  A. Sharrocks,et al.  MADS-box transcription factors adopt alternative mechanisms for bending DNA. , 1999, Journal of molecular biology.

[36]  Clarence S. M. Chan,et al.  Organization of DNA sequences and replication origins at yeast telomeres , 1983, Cell.

[37]  D. Liberles,et al.  Role of architectural elements in combinatorial regulation of initiation of DNA replication in Escherichia coli , 1997, Molecular microbiology.

[38]  D. Helinski,et al.  A Critical DnaA Box Directs the Cooperative Binding of theEscherichia coli DnaA Protein to the Plasmid RK2 Replication Origin* , 1999, The Journal of Biological Chemistry.

[39]  B. Tye,et al.  Autonomously replicating sequences in Saccharomyces cerevisiae. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[40]  E. Kremmer,et al.  Human origin recognition complex binds to the region of the latent origin of DNA replication of Epstein–Barr virus , 2001, The EMBO journal.

[41]  J. Blow,et al.  Replication Origins in XenopusEgg Extract Are 5–15 Kilobases Apart and Are Activated in Clusters That Fire at Different Times , 2001, The Journal of cell biology.

[42]  R. Elble,et al.  Saccharomyces cerevisiae protein involved in plasmid maintenance is necessary for mating of MAT alpha cells. , 1988, Journal of molecular biology.

[43]  H. Xu,et al.  Human DNA replication initiation factors, ORC and MCM, associate with oriP of Epstein–Barr virus , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[44]  Bruce Stillman,et al.  ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex , 1992, Nature.

[45]  S. Eisenberg,et al.  Analysis of the interactions of functional domains of a nuclear origin of replication from Saccharomyces cerevisiae. , 1991, Nucleic acids research.

[46]  S. Gerbi,et al.  Chromosomal ARS1 has a single leading strand start site. , 1999, Molecular cell.

[47]  C. Newlon,et al.  Evidence suggesting that the ARS elements associated with silencers of the yeast mating-type locus HML do not function as chromosomal DNA replication origins , 1991, Molecular and cellular biology.

[48]  Ronald W. Davis,et al.  Replication dynamics of the yeast genome. , 2001, Science.

[49]  S. Bell,et al.  Nucleosomes positioned by ORC facilitate the initiation of DNA replication. , 2001, Molecular cell.

[50]  M. Kladde,et al.  Tup1p represses Mcm1p transcriptional activation and chromatin remodeling of an a‐cell‐specific gene , 2000, The EMBO journal.

[51]  J. Diffley,et al.  Purification of a yeast protein that binds to origins of DNA replication and a transcriptional silencer. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[52]  Lukas Endler,et al.  Forkhead-like transcription factors recruit Ndd1 to the chromatin of G2/M-specific promoters , 2000, Nature.

[53]  T. Kelly,et al.  Regulation of chromosome replication. , 2000, Annual review of biochemistry.

[54]  C. Newlon,et al.  Domain B of ARS307 contains two functional elements and contributes to chromosomal replication origin function , 1994, Molecular and cellular biology.

[55]  W. L. Fangman,et al.  The localization of replication origins on ARS plasmids in S. cerevisiae , 1987, Cell.