This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Dalton, S.
Right arrow Articles by Hopwood, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Dalton, S.
Right arrow Articles by Hopwood, B.

 Previous Article  |  Next Article 

Mol. Cell. Biol., 10 1997, 5867-5875, Vol 17, No. 10
Copyright © 1997, American Society for Microbiology

Characterization of Cdc47p-minichromosome maintenance complexes in Saccharomyces cerevisiae: identification of Cdc45p as a subunit

S Dalton and B Hopwood
Department of Biochemistry, University of Adelaide, South Australia. sdalton@biochem.adelaide.edu.au

Cdc47p is a member of the minichromosome maintenance (MCM) family of polypeptides, which have a role in the early stages of chromosomal DNA replication. Here, we show that Cdc47p assembles into stable complexes with two other members of the MCM family, Cdc46p and Mcm3p. The assembly of Cdc47p into complexes with Cdc46p does not appear to be cell cycle regulated, making it unlikely that these interactions per se are a rate-limiting step in the control of S phase. Cdc45p is also shown to interact with Cdc47p in vivo and to be a component of high- molecular-weight MCM complexes in cell lysates. Like MCM polypeptides, Cdc45p is essential for the initiation of chromosomal DNA replication in Saccharomyces cerevisiae; however, Cdc45p remains in the nucleus throughout the cell cycle, whereas MCMs are nuclear only during G1. We characterize two mutations in CDC47 and CDC46 which arrest cells with unduplicated DNA as a result of single base substitutions. The corresponding amino acid substitutions in Cdc46p and Cdc47p severely reduce the ability of these polypeptides to assemble in a complex with each other in vivo and in vitro. This argues that assembly of Cdc47p into complexes with other MCM polypeptides is important for its role in the initiation of chromosomal DNA replication.


This article has been cited by other articles:

  • Leon, R. P., Tecklenburg, M., Sclafani, R. A. (2008). Functional Conservation of {beta}-Hairpin DNA Binding Domains in the Mcm Protein of Methanobacterium thermoautotrophicum and the Mcm5 protein of Saccharomyces cerevisiae. Genetics 179: 1757-1768 [Abstract] [Full Text]  
  • Forsburg, S. L. (2004). Eukaryotic MCM Proteins: Beyond Replication Initiation. Microbiol. Mol. Biol. Rev. 68: 109-131 [Abstract] [Full Text]  
  • Davey, M. J., Indiani, C., O'Donnell, M. (2003). Reconstitution of the Mcm2-7p Heterohexamer, Subunit Arrangement, and ATP Site Architecture. J. Biol. Chem. 278: 4491-4499 [Abstract] [Full Text]  
  • Schaarschmidt, D., Ladenburger, E.-M., Keller, C., Knippers, R. (2002). Human Mcm proteins at a replication origin during the G1 to S phase transition. Nucleic Acids Res 30: 4176-4185 [Abstract] [Full Text]  
  • Nakajima, R., Masukata, H. (2002). SpSld3 Is Required for Loading and Maintenance of SpCdc45 on Chromatin in DNA Replication in Fission Yeast. Mol. Biol. Cell 13: 1462-1472 [Abstract] [Full Text]  
  • Liang, D. T., Forsburg, S. L. (2001). Characterization of Schizosaccharomyces pombe mcm7+ and cdc23+ (MCM10) and Interactions With Replication Checkpoints. Genetics 159: 471-486 [Abstract] [Full Text]  
  • Yoshida, K., Kuo, F., George, E. L., Sharpe, A. H., Dutta, A. (2001). Requirement of CDC45 for Postimplantation Mouse Development. Mol. Cell. Biol. 21: 4598-4603 [Abstract] [Full Text]  
  • Loebel, D., Huikeshoven, H., Cotterill, S. (2000). Localisation of the DmCdc45 DNA replication factor in the mitotic cycle and during chorion gene amplification. Nucleic Acids Res 28: 3897-3903 [Abstract] [Full Text]  
  • Zou, L., Stillman, B. (2000). Assembly of a Complex Containing Cdc45p, Replication Protein A, and Mcm2p at Replication Origins Controlled by S-Phase Cyclin-Dependent Kinases and Cdc7p-Dbf4p Kinase. Mol. Cell. Biol. 20: 3086-3096 [Abstract] [Full Text]  
  • Masumoto, H., Sugino, A., Araki, H. (2000). Dpb11 Controls the Association between DNA Polymerases alpha and varepsilon and the Autonomously Replicating Sequence Region of Budding Yeast. Mol. Cell. Biol. 20: 2809-2817 [Abstract] [Full Text]  
  • Homesley, L., Lei, M., Kawasaki, Y., Sawyer, S., Christensen, T., Tye, B. K. (2000). Mcm10 and the MCM2-7 complex interact to initiate DNA synthesis and to release replication factors from origins. Genes Dev. 14: 913-926 [Abstract] [Full Text]  
  • Arata, Y., Fujita, M., Ohtani, K., Kijima, S., Kato, J.-y. (2000). Cdk2-dependent and -independent Pathways in E2F-mediated S Phase Induction. J. Biol. Chem. 275: 6337-6345 [Abstract] [Full Text]  
  • Springer, P., Holding, D., Groover, A, Yordan, C, Martienssen, R. (2000). The essential Mcm7 protein PROLIFERA is localized to the nucleus of dividing cells during the G(1) phase and is required maternally for early Arabidopsis development. Development 127: 1815-1822 [Abstract]  
  • Reid, R. J. D., Fiorani, P., Sugawara, M., Bjornsti, M.-A. (1999). CDC45 and DPB11 are required for processive DNA replication and resistance to DNA topoisomerase I-mediated DNA damage. Proc. Natl. Acad. Sci. USA 96: 11440-11445 [Abstract] [Full Text]  
  • Oshiro, G., Owens, J. C., Shellman, Y., Sclafani, R. A., Li, J. J. (1999). Cell Cycle Control of Cdc7p Kinase Activity through Regulation of Dbf4p Stability. Mol. Cell. Biol. 19: 4888-4896 [Abstract] [Full Text]  
  • Kamimura, Y., Masumoto, H., Sugino, A., Araki, H. (1998). Sld2, Which Interacts with Dpb11 in Saccharomyces cerevisiae, Is Required for Chromosomal DNA Replication. Mol. Cell. Biol. 18: 6102-6109 [Abstract] [Full Text]  
  • Sherman, D. A., Pasion, S. G., Forsburg, S. L. (1998). Multiple Domains of Fission Yeast Cdc19p (MCM2) Are Required for Its Association with the Core MCM Complex. Mol. Biol. Cell 9: 1833-1845 [Abstract] [Full Text]  
  • Zou, L., Stillman, B. (1998). Formation of a Preinitiation Complex by S-phase Cyclin CDK-Dependent Loading of Cdc45p onto Chromatin. Science 280: 593-596 [Abstract] [Full Text]  
  • Lee, J.-K., Hurwitz, J. (2000). Isolation and Characterization of Various Complexes of the Minichromosome Maintenance Proteins of Schizosaccharomyces pombe. J. Biol. Chem. 275: 18871-18878 [Abstract] [Full Text]  
  • Knoop, L. L., Baker, S. J. (2000). The Splicing Factor U1C Represses EWS/FLI-mediated Transactivation. J. Biol. Chem. 275: 24865-24871 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»