Mammalian DNA Polymerase Auxiliary Proteins: Analysis of Replication Factor C-catalyzed Proliferating Cell Nuclear Antigen Loading Onto Circular Double-stranded DNA

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 1995 Jun 1

PMID 7760803

Citations 37

Authors

L M Podust

V N Podust

J M Sogo

U Hubscher

Affiliations

Soon will be listed here.

Abstract

To understand the mechanism of action of the two eukaryotic replication auxiliary proteins proliferating cell nuclear antigen (PCNA) and replication factor C (RF-C), we constructed a plasmid for producing PCNA which could be 32P labelled in vitro. This allowed us to analyze the assembly of the auxiliary proteins directly on DNA and to examine this process in the absence of DNA synthesis. By using closed circular double-stranded DNA or gapped circular DNA for protein-DNA complex formation, the following results were obtained, (i) RF-C can load PCNA in an ATP-dependent manner directly on double-stranded DNA, and no 3'-OH ends are required for this reaction; (ii) the RF-C-PCNA complex assembled on closed circular DNA differs from those assembled on gapped or nicked circular DNA; (iii) the stable RF-C-PCNA complex can be assembled on circular but not on linear DNA; and (iv) only gapped DNA can partially retain the assembled RF-C-PCNA complex upon the linearization of the template. We propose that RF-C first binds unspecifically to double-stranded DNA in the presence of ATP and then loads PCNA onto DNA to yield a protein complex able to track along DNA. The RF-C-PCNA complex could slide along the template until it encounters a 3'-OH primer-template junction, where it is likely transformed into a competent clamp. The latter complex, finally, might still be able to slide along double-stranded DNA.

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References

Stillman B . Smart machines at the DNA replication fork. Cell. 1994; 78(5):725-8. DOI: 10.1016/s0092-8674(94)90362-x. View

Tsurimoto T, Stillman B . Replication factors required for SV40 DNA replication in vitro. II. Switching of DNA polymerase alpha and delta during initiation of leading and lagging strand synthesis. J Biol Chem. 1991; 266(3):1961-8. View

Weiser T, Gassmann M, Thommes P, Ferrari E, Hafkemeyer P, Hubscher U . Biochemical and functional comparison of DNA polymerases alpha, delta, and epsilon from calf thymus. J Biol Chem. 1991; 266(16):10420-8. View

Stukenberg P, ODonnell M . Mechanism of the sliding beta-clamp of DNA polymerase III holoenzyme. J Biol Chem. 1991; 266(17):11328-34. View

Burgers P . Saccharomyces cerevisiae replication factor C. II. Formation and activity of complexes with the proliferating cell nuclear antigen and with DNA polymerases delta and epsilon. J Biol Chem. 1991; 266(33):22698-706. View

Lee S, Pan Z, Kwong A, Burgers P, Hurwitz J . Synthesis of DNA by DNA polymerase epsilon in vitro. J Biol Chem. 1991; 266(33):22707-17. View

Fien K, Stillman B . Identification of replication factor C from Saccharomyces cerevisiae: a component of the leading-strand DNA replication complex. Mol Cell Biol. 1992; 12(1):155-63. PMC: 364079. DOI: 10.1128/mcb.12.1.155-163.1992. View

Gogol E, Young M, Kubasek W, Jarvis T, von Hippel P . Cryoelectron microscopic visualization of functional subassemblies of the bacteriophage T4 DNA replication complex. J Mol Biol. 1992; 224(2):395-412. DOI: 10.1016/0022-2836(92)91003-8. View

Kong X, Onrust R, ODonnell M, Kuriyan J . Three-dimensional structure of the beta subunit of E. coli DNA polymerase III holoenzyme: a sliding DNA clamp. Cell. 1992; 69(3):425-37. DOI: 10.1016/0092-8674(92)90445-i. View

10.

Podust V, Georgaki A, Strack B, Hubscher U . Calf thymus RF-C as an essential component for DNA polymerase delta and epsilon holoenzymes function. Nucleic Acids Res. 1992; 20(16):4159-65. PMC: 334120. DOI: 10.1093/nar/20.16.4159. View

11.

Pan Z, Chen M, Hurwitz J . The subunits of activator 1 (replication factor C) carry out multiple functions essential for proliferating-cell nuclear antigen-dependent DNA synthesis. Proc Natl Acad Sci U S A. 1993; 90(1):6-10. PMC: 45588. DOI: 10.1073/pnas.90.1.6. View

12.

ODonnell M, Onrust R, Dean F, Chen M, Hurwitz J . Homology in accessory proteins of replicative polymerases--E. coli to humans. Nucleic Acids Res. 1993; 21(1):1-3. PMC: 309057. DOI: 10.1093/nar/21.1.1. View

13.

Podust V, Hubscher U . Lagging strand DNA synthesis by calf thymus DNA polymerases alpha, beta, delta and epsilon in the presence of auxiliary proteins. Nucleic Acids Res. 1993; 21(4):841-6. PMC: 309215. DOI: 10.1093/nar/21.4.841. View

14.

Lu Y, Zeft A, Riegel A . Cloning and expression of a novel human DNA binding protein, PO-GA. Biochem Biophys Res Commun. 1993; 193(2):779-86. DOI: 10.1006/bbrc.1993.1693. View

15.

Burgers P, Yoder B . ATP-independent loading of the proliferating cell nuclear antigen requires DNA ends. J Biol Chem. 1993; 268(27):19923-6. View

16.

Kaboord B, Benkovic S . Rapid assembly of the bacteriophage T4 core replication complex on a linear primer/template construct. Proc Natl Acad Sci U S A. 1993; 90(22):10881-5. PMC: 47882. DOI: 10.1073/pnas.90.22.10881. View

17.

Bunz F, Kobayashi R, Stillman B . cDNAs encoding the large subunit of human replication factor C. Proc Natl Acad Sci U S A. 1993; 90(23):11014-8. PMC: 47912. DOI: 10.1073/pnas.90.23.11014. View

18.

Kuriyan J, ODonnell M . Sliding clamps of DNA polymerases. J Mol Biol. 1993; 234(4):915-25. DOI: 10.1006/jmbi.1993.1644. View

19.

Burbelo P, Utani A, Pan Z, Yamada Y . Cloning of the large subunit of activator 1 (replication factor C) reveals homology with bacterial DNA ligases. Proc Natl Acad Sci U S A. 1993; 90(24):11543-7. PMC: 48020. DOI: 10.1073/pnas.90.24.11543. View

20.

Luckow B, Bunz F, Stillman B, Lichter P, Schutz G . Cloning, expression, and chromosomal localization of the 140-kilodalton subunit of replication factor C from mice and humans. Mol Cell Biol. 1994; 14(3):1626-34. PMC: 358521. DOI: 10.1128/mcb.14.3.1626-1634.1994. View