Mcm2 is a Target of Regulation by Cdc7-Dbf4 During the Initiation of DNA Synthesis

Overview

Journal Genes Dev

Specialty Molecular Biology

Date 1998 Feb 7

PMID 9407029

Citations 139

Authors

M Lei

Y Kawasaki

M R Young

M Kihara

A Sugino

B K Tye

Affiliations

Soon will be listed here.

Abstract

The initiation of DNA synthesis is an important cell cycle event that defines the beginning of S phase. This critical event involves the participation of proteins whose functions are regulated by cyclin dependent protein kinases (Cdks). The Mcm2-7 proteins are a family of six conserved proteins that are essential for the initiation of DNA synthesis in all eukaryotes. In Saccharomyces cerevisiae, members of the Mcm2-7 family undergo cell cycle-specific phosphorylation. Phosphorylation of Mcm proteins at the beginning of S phase coincides with the removal of these proteins from chromatin and the onset of DNA synthesis. In this study, we identified DBF4, which encodes the regulatory subunit of a Cdk-like protein kinase Cdc7-Dbf4, in a screen for second site suppressors of mcm2-1. The dbf4 suppressor mutation restores competence to initiate DNA synthesis to the mcm2-1 mutant. Cdc7-Dbf4 interacts physically with Mcm2 and phosphorylates Mcm2 and three other members of the Mcm2-7 family in vitro. Blocking the kinase activity of Cdc7-Dbf4 at the G1-to-S phase transition also blocks the phosphorylation of Mcm2 at this defined point of the cell cycle. Taken together, our data suggest that phosphorylation of Mcm2 and probably other members of the Mcm2-7 proteins by Cdc7-Dbf4 at the G1-to-S phase transition is a critical step in the initiation of DNA synthesis at replication origins.

Citing Articles

Development of a novel centrosome-related risk signature to predict prognosis and treatment response in lung adenocarcinoma.

Wang Z, Zuo C, Fei J, Chen H, Wang L, Xie Y Discov Oncol. 2024; 15(1):717.

PMID: 39592523 PMC: 11599701. DOI: 10.1007/s12672-024-01615-8.

Four decades of Eukaryotic DNA replication: From yeast genetics to high-resolution cryo-EM structures of the replisome.

Tye B Proc Natl Acad Sci U S A. 2024; 121(42):e2415231121.

PMID: 39365830 PMC: 11494305. DOI: 10.1073/pnas.2415231121.

Assembly, Activation, and Helicase Actions of MCM2-7: Transition from Inactive MCM2-7 Double Hexamers to Active Replication Forks.

You Z, Masai H Biology (Basel). 2024; 13(8).

PMID: 39194567 PMC: 11351656. DOI: 10.3390/biology13080629.

RUNX1 regulates MCM2/CDC20 to promote COAD progression modified by deubiquitination of USP31.

Tian W, Zhao J, Zhang X, Li P, Li X, Hong Y Sci Rep. 2024; 14(1):13906.

PMID: 38886545 PMC: 11183096. DOI: 10.1038/s41598-024-64726-w.

Netrin 1 directs vascular patterning and maturity in the developing kidney.

Honeycutt S, NGuetta P, Hardesty D, Xiong Y, Cooper S, Stevenson M Development. 2023; 150(22).

PMID: 37818607 PMC: 10690109. DOI: 10.1242/dev.201886.

References

Adams A, Botstein D, Drubin D . A yeast actin-binding protein is encoded by SAC6, a gene found by suppression of an actin mutation. Science. 1989; 243(4888):231-3. DOI: 10.1126/science.2643162. View

Young M, Suzuki K, Yan H, Gibson S, Tye B . Nuclear accumulation of Saccharomyces cerevisiae Mcm3 is dependent on its nuclear localization sequence. Genes Cells. 1998; 2(10):631-43. DOI: 10.1046/j.1365-2443.1997.1510349.x. View

Coue M, Kearsey S, Mechali M . Chromatin binding, nuclear localization and phosphorylation of Xenopus cdc21 are cell-cycle dependent and associated with the control of initiation of DNA replication. EMBO J. 1996; 15(5):1085-97. PMC: 450006. View

LEATHERWOOD J, Lopez-Girona A, Russell P . Interaction of Cdc2 and Cdc18 with a fission yeast ORC2-like protein. Nature. 1996; 379(6563):360-3. DOI: 10.1038/379360a0. View

Stillman B . Cell cycle control of DNA replication. Science. 1996; 274(5293):1659-64. DOI: 10.1126/science.274.5293.1659. View

Merchant A, Kawasaki Y, Chen Y, Lei M, Tye B . A lesion in the DNA replication initiation factor Mcm10 induces pausing of elongation forks through chromosomal replication origins in Saccharomyces cerevisiae. Mol Cell Biol. 1997; 17(6):3261-71. PMC: 232179. DOI: 10.1128/MCB.17.6.3261. View

Kitada K, Johnson A, Johnston L, Sugino A . A multicopy suppressor gene of the Saccharomyces cerevisiae G1 cell cycle mutant gene dbf4 encodes a protein kinase and is identified as CDC5. Mol Cell Biol. 1993; 13(7):4445-57. PMC: 360015. DOI: 10.1128/mcb.13.7.4445-4457.1993. View

Chong J, Mahbubani H, Khoo C, J Blow J . Purification of an MCM-containing complex as a component of the DNA replication licensing system. Nature. 1995; 375(6530):418-21. DOI: 10.1038/375418a0. View

Cocker J, Piatti S, Santocanale C, Nasmyth K, Diffley J . An essential role for the Cdc6 protein in forming the pre-replicative complexes of budding yeast. Nature. 1996; 379(6561):180-2. DOI: 10.1038/379180a0. View

10.

Yan H, Merchant A, Tye B . 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; 7(11):2149-60. DOI: 10.1101/gad.7.11.2149. View

11.

Kearsey S, Maiorano D, Holmes E, Todorov I . The role of MCM proteins in the cell cycle control of genome duplication. Bioessays. 1996; 18(3):183-90. DOI: 10.1002/bies.950180305. View

12.

Brewer B, FANGMAN W . The localization of replication origins on ARS plasmids in S. cerevisiae. Cell. 1987; 51(3):463-71. DOI: 10.1016/0092-8674(87)90642-8. View

13.

Aparicio O, Weinstein D, Bell S . Components and dynamics of DNA replication complexes in S. cerevisiae: redistribution of MCM proteins and Cdc45p during S phase. Cell. 1997; 91(1):59-69. DOI: 10.1016/s0092-8674(01)80009-x. View

14.

Thommes P, Kubota Y, Takisawa H, J Blow J . The RLF-M component of the replication licensing system forms complexes containing all six MCM/P1 polypeptides. EMBO J. 1997; 16(11):3312-9. PMC: 1169947. DOI: 10.1093/emboj/16.11.3312. View

15.

Bell S, Stillman B . ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex. Nature. 1992; 357(6374):128-34. DOI: 10.1038/357128a0. View

16.

Kitada K, Johnston L, Sugino T, Sugino A . Temperature-sensitive cdc7 mutations of Saccharomyces cerevisiae are suppressed by the DBF4 gene, which is required for the G1/S cell cycle transition. Genetics. 1992; 131(1):21-9. PMC: 1204955. DOI: 10.1093/genetics/131.1.21. View

17.

Schild D, Byers B . Meiotic effects of DNA-defective cell division cycle mutations of Saccharomyces cerevisiae. Chromosoma. 1978; 70(1):109-30. DOI: 10.1007/BF00292220. View

18.

Hutter K, Eipel H . Flow cytometric determinations of cellular substances in algae, bacteria, moulds and yeasts. Antonie Van Leeuwenhoek. 1978; 44(3-4):269-82. DOI: 10.1007/BF00394305. View

19.

Diffley J . Once and only once upon a time: specifying and regulating origins of DNA replication in eukaryotic cells. Genes Dev. 1996; 10(22):2819-30. DOI: 10.1101/gad.10.22.2819. View

20.

Madine M, Khoo C, Mills A, Laskey R . MCM3 complex required for cell cycle regulation of DNA replication in vertebrate cells. Nature. 1995; 375(6530):421-4. DOI: 10.1038/375421a0. View