Home LiteratureArticle Details
PMID: 9407030 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Persistent initiation of DNA replication and chromatin-bound MCM proteins during the cell cycle in cdc6 mutants.

Genes & development ·Vol. 11 ·No. 24 ·1997-12-15 ·Pages 3375-86

Liang C, Stillman B

Abstract

Faithful inheritance of genetic information requires that DNA be copied only once each cell cycle. Initiation of DNA replication involves the establishment of a prereplication complex (pre-RC) and subsequent activation by CDK/cyclins, converting the pre-RC to a post-RC. The origin recognition complex (ORC), Cdc6p, and the MCM proteins are required for establishing the pre-RC. We show that all six ORC subunits remain bound to chromatin throughout the cell cycle, whereas the MCM proteins cycle on and off, corresponding precisely to transitions of the RC. A newly isolated cdc6 mutant displays promiscuous initiation of DNA replication, increased nuclear DNA content, and constant MCM protein association with chromatin throughout the cell cycle. This gain-of-function cdc6 mutant ignores the negative controls imposed normally on initiation by the CDK/cyclins, suggesting that Cdc6p is a key mediator of once-per-cell-cycle control of DNA replication.

MeSH Terms
CDC28 Protein Kinase, S cerevisiae/metabolism Cell Cycle/genetics Cell Cycle Proteins/genetics,metabolism Chromatin/metabolism Chromosomal Proteins, Non-Histone DNA Replication Fungal Proteins/genetics,metabolism Mutation Replication Origin Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins Temperature
Chemicals
CDC6 protein, S cerevisiae Cell Cycle Proteins Chromatin Chromosomal Proteins, Non-Histone Fungal Proteins Saccharomyces cerevisiae Proteins CDC28 Protein Kinase, S cerevisiae MCM2 protein, S cerevisiae
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Liang C
Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724 USA.
Stillman B
References (48)
48 references, click to expand
  1. The origin recognition complex in silencing, cell cycle progression, and DNA replication.
    Mol Biol Cell. 1995 Jun;6(6):741-56 PMID: 7579692
  2. Cdc6 is an unstable protein whose de novo synthesis in G1 is important for the onset of S phase and for preventing a 'reductional' anaphase in the budding yeast Saccharomyces cerevisiae.
    EMBO J. 1995 Aug 1;14(15):3788-99 PMID: 7641697
  3. The phenotype of the minichromosome maintenance mutant mcm3 is characteristic of mutants defective in DNA replication.
    Mol Cell Biol. 1990 Nov;10(11):5707-20 PMID: 2233713
  4. CDC6 mRNA fluctuates periodically in the yeast cell cycle.
    J Biol Chem. 1990 Nov 15;265(32):19904-9 PMID: 2246267
  5. A group of interacting yeast DNA replication genes.
    Genes Dev. 1991 Jun;5(6):958-69 PMID: 2044962
  6. ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex.
    Nature. 1992 May 14;357(6374):128-34 PMID: 1579162
  7. Protein-DNA interactions at a yeast replication origin.
    Nature. 1992 May 14;357(6374):169-72 PMID: 1579168
  8. Dual functions of CDC6: a yeast protein required for DNA replication also inhibits nuclear division.
    EMBO J. 1992 Jun;11(6):2167-76 PMID: 1600944
  9. In vitro reactions of vacuole inheritance in Saccharomyces cerevisiae.
    J Cell Biol. 1992 Dec;119(6):1469-79 PMID: 1334958
  10. Cell cycle-regulated nuclear localization of MCM2 and MCM3, which are required for the initiation of DNA synthesis at chromosomal replication origins in yeast.
    Genes Dev. 1993 Nov;7(11):2149-60 PMID: 8224843
  11. Yeast origin recognition complex functions in transcription silencing and DNA replication.
    Science. 1993 Dec 17;262(5141):1844-9 PMID: 8266072
  12. The initiator protein DnaA: evolution, properties and function.
    Biochim Biophys Acta. 1994 Mar 1;1217(2):111-30 PMID: 8110826
  13. Hyperactive initiation of chromosomal replication in vivo and in vitro by a mutant initiator protein, DnaAcos, of Escherichia coli.
    J Biol Chem. 1994 Apr 29;269(17):12698-703 PMID: 8175680
  14. Two steps in the assembly of complexes at yeast replication origins in vivo.
    Cell. 1994 Jul 29;78(2):303-16 PMID: 8044842
  15. The mutant DnaAcos protein which overinitiates replication of the Escherichia coli chromosome is inert to negative regulation for initiation.
    J Biol Chem. 1994 Sep 2;269(35):22075-9 PMID: 8071330
  16. The Saccharomyces cerevisiae CDC6 gene is transcribed at late mitosis and encodes a ATP/GTPase controlling S phase initiation.
    J Biol Chem. 1994 Sep 16;269(37):23351-6 PMID: 8083240
  17. The multidomain structure of Orc1p reveals similarity to regulators of DNA replication and transcriptional silencing.
    Cell. 1995 Nov 17;83(4):563-8 PMID: 7585959
  18. An essential role for the Cdc6 protein in forming the pre-replicative complexes of budding yeast.
    Nature. 1996 Jan 11;379(6561):180-2 PMID: 8538771
  19. S-phase-promoting cyclin-dependent kinases prevent re-replication by inhibiting the transition of replication origins to a pre-replicative state.
    Curr Biol. 1995 Nov 1;5(11):1257-69 PMID: 8574583
  20. The yeast CDC16 and CDC27 genes restrict DNA replication to once per cell cycle.
    Cell. 1996 Apr 5;85(1):39-48 PMID: 8620535
  21. cdc18+ regulates initiation of DNA replication in Schizosaccharomyces pombe.
    Proc Natl Acad Sci U S A. 1996 Feb 20;93(4):1566-70 PMID: 8643672
  22. Coordinating DNA replication to produce one copy of the genome requires genes that act in ubiquitin metabolism.
    Mol Cell Biol. 1996 Apr;16(4):1356-66 PMID: 8657109
  23. Activation of S-phase-promoting CDKs in late G1 defines a "point of no return" after which Cdc6 synthesis cannot promote DNA replication in yeast.
    Genes Dev. 1996 Jun 15;10(12):1516-31 PMID: 8666235
  24. pMPY-ZAP: a reusable polymerase chain reaction-directed gene disruption cassette for Saccharomyces cerevisiae.
    Yeast. 1996 Feb;12(2):129-34 PMID: 8686376
  25. Physical interactions among Mcm proteins and effects of Mcm dosage on DNA replication in Saccharomyces cerevisiae.
    Mol Cell Biol. 1996 Sep;16(9):5081-90 PMID: 8756666
  26. The Xenopus Cdc6 protein is essential for the initiation of a single round of DNA replication in cell-free extracts.
    Cell. 1996 Oct 4;87(1):53-63 PMID: 8858148
  27. The role of MCM/P1 proteins in the licensing of DNA replication.
    Trends Biochem Sci. 1996 Mar;21(3):102-6 PMID: 8882583
  28. Once and only once upon a time: specifying and regulating origins of DNA replication in eukaryotic cells.
    Genes Dev. 1996 Nov 15;10(22):2819-30 PMID: 8918884
  29. Interaction between yeast Cdc6 protein and B-type cyclin/Cdc28 kinases.
    Mol Biol Cell. 1996 Nov;7(11):1723-35 PMID: 8930895
  30. Cell cycle control of DNA replication.
    Science. 1996 Dec 6;274(5293):1659-64 PMID: 8939847
  31. Viewpoint: putting the cell cycle in order.
    Science. 1996 Dec 6;274(5293):1643-5 PMID: 8984634
  32. Coordinate binding of ATP and origin DNA regulates the ATPase activity of the origin recognition complex.
    Cell. 1997 Feb 21;88(4):493-502 PMID: 9038340
  33. Cdc6p establishes and maintains a state of replication competence during G1 phase.
    J Cell Sci. 1997 Mar;110 ( Pt 6):753-63 PMID: 9099949
  34. A role for Cdk2 kinase in negatively regulating DNA replication during S phase of the cell cycle.
    J Cell Biol. 1997 Apr 7;137(1):183-92 PMID: 9105046
  35. The k43 gene, required for chorion gene amplification and diploid cell chromosome replication, encodes the Drosophila homolog of yeast origin recognition complex subunit 2.
    Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3888-92 PMID: 9108074
  36. Cyclin-dependent kinase and initiation at eukaryotic origins: a replication switch?
    Curr Opin Cell Biol. 1997 Jun;9(3):358-63 PMID: 9159077
  37. Cdc6p-dependent loading of Mcm proteins onto pre-replicative chromatin in budding yeast.
    Proc Natl Acad Sci U S A. 1997 May 27;94(11):5611-6 PMID: 9159120
  38. Loading of an Mcm protein onto DNA replication origins is regulated by Cdc6p and CDKs.
    Cell. 1997 Aug 22;90(4):649-60 PMID: 9288745
  39. The B-type cyclin kinase inhibitor p40SIC1 controls the G1 to S transition in S. cerevisiae.
    Cell. 1994 Oct 21;79(2):233-44 PMID: 7954792
  40. Cell cycle-regulated nuclear import and export of Cdc47, a protein essential for initiation of DNA replication in budding yeast.
    Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2514-8 PMID: 7708676
  41. Identification of the yeast MCM3-related protein as a component of Xenopus DNA replication licensing factor.
    Cell. 1995 May 19;81(4):601-9 PMID: 7758114
  42. Purification of an MCM-containing complex as a component of the DNA replication licensing system.
    Nature. 1995 Jun 1;375(6530):418-21 PMID: 7760937
  43. MCM3 complex required for cell cycle regulation of DNA replication in vertebrate cells.
    Nature. 1995 Jun 1;375(6530):421-4 PMID: 7760938
  44. ORC and Cdc6p interact and determine the frequency of initiation of DNA replication in the genome.
    Cell. 1995 Jun 2;81(5):667-76 PMID: 7774008
  45. The origin recognition complex has essential functions in transcriptional silencing and chromosomal replication.
    Genes Dev. 1995 Apr 15;9(8):911-24 PMID: 7774809
  46. BM28, a human member of the MCM2-3-5 family, is displaced from chromatin during DNA replication.
    J Cell Biol. 1995 Jun;129(6):1433-45 PMID: 7790346
  47. Molecular cloning of cDNA encoding mouse Cdc21 and CDC46 homologs and characterization of the products: physical interaction between P1(MCM3) and CDC46 proteins.
    Nucleic Acids Res. 1995 Jun 25;23(12):2097-104 PMID: 7610039
  48. p65cdc18 plays a major role controlling the initiation of DNA replication in fission yeast.
    Cell. 1995 Nov 3;83(3):397-405 PMID: 8521469
Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.11.24.3375
Published
1997-12-15
Pages
3375-86
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC316796
Subset
IM
Grants
NIGMS NIH HHS · GM45436 · United States
External Links
PubMed source DOI: 10.1101/gad.11.24.3375 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2