Home LiteratureArticle Details
PMID: 1363314 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

DNA polymerase delta and epsilon holoenzymes from calf thymus.

Chromosoma ·Vol. 102 ·No. 1 Suppl ·1992-00-00 ·Pages S133-41

Podust V, Mikhailov V, Georgaki A, Hübscher U

Abstract

Replication of singly-DNA primed M13 DNA by DNA polymerase (pol) delta completely relies on the simultaneous addition of proliferating cell nuclear antigen (PCNA), replication factor C (RF-C) and replication protein A (RP-A) (or E. coli single-strand DNA binding protein, SSB). Pol epsilon core alone is able to synthesize the products on singly-primed ssDNA. However, DNA synthesis by pol epsilon was stimulated up to 10-fold upon addition of the three auxiliary proteins PCNA, RF-C and SSB. This stimulation of pol epsilon by PCNA/RF-C/SSB appears to be the superposition of two events: pol epsilon holoenzyme (pol epsilon, PCNA, RF-C) synthesized longer products than its pol epsilon core counterpart, but elongated less primers. Furthermore, we analyzed the cooperative action of pol alpha/primase with pol delta or pol epsilon holoenzymes on unprimed M13 DNA. While pol delta displayed higher dNMP incorporation than pol epsilon, when a single primer was preannealed to DNA, pol epsilon was more efficient in the utilization of the primers synthesized by pol alpha/primase. Under these conditions both longer products and a higher amount of dNMP incorporation was found for pol epsilon holoenzyme, than for pol delta. Our data support the hypothesis of pol delta as the leading and pol epsilon as the second lagging strand replication enzyme.

MeSH Terms
Animals Cattle DNA Polymerase II/physiology DNA Polymerase III DNA Primase DNA Replication/physiology DNA, Single-Stranded/metabolism DNA-Binding Proteins/physiology DNA-Directed DNA Polymerase/metabolism Homeodomain Proteins Minor Histocompatibility Antigens Nuclear Proteins/physiology Proliferating Cell Nuclear Antigen Proto-Oncogene Proteins c-bcl-2 RNA Nucleotidyltransferases/physiology Replication Protein C Repressor Proteins Saccharomyces cerevisiae Proteins Thymus Gland/enzymology
Chemicals
BCL2-related protein A1 DNA, Single-Stranded DNA-Binding Proteins Homeodomain Proteins MATA1 protein, S cerevisiae Minor Histocompatibility Antigens Nuclear Proteins Proliferating Cell Nuclear Antigen Proto-Oncogene Proteins c-bcl-2 Repressor Proteins Saccharomyces cerevisiae Proteins DNA Primase RNA Nucleotidyltransferases DNA Polymerase II DNA Polymerase III DNA-Directed DNA Polymerase Replication Protein C
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Podust V
Department of Pharmacology and Biochemistry, University Zürich-Irchel, Switzerland.
Mikhailov V
Georgaki A
Hübscher U
References (34)
34 references, click to expand
  1. Functions of replication factor C and proliferating-cell nuclear antigen: functional similarity of DNA polymerase accessory proteins from human cells and bacteriophage T4.
    Proc Natl Acad Sci U S A. 1990 Feb;87(3):1023-7 PMID: 1967833
  2. DNA polymerase III holoenzyme. Components, structure, and mechanism of a true replicative complex.
    J Biol Chem. 1991 Oct 15;266(29):19127-30 PMID: 1918028
  3. Calf thymus DNA polymerase delta independent of proliferating cell nuclear antigen (PCNA).
    Nucleic Acids Res. 1989 Mar 11;17(5):1805-21 PMID: 2564661
  4. Two DNA polymerases may be required for synthesis of the lagging DNA strand of simian virus 40.
    J Virol. 1990 Dec;64(12):5912-8 PMID: 2173773
  5. Studies on the activator 1 protein complex, an accessory factor for proliferating cell nuclear antigen-dependent DNA polymerase delta.
    J Biol Chem. 1991 Jan 5;266(1):594-602 PMID: 1670772
  6. Studies of the cloned 37-kDa subunit of activator 1 (replication factor C) of HeLa cells.
    Proc Natl Acad Sci U S A. 1992 Jun 15;89(12 ):5211-5 PMID: 1351677
  7. Binding properties of replication protein A from human and yeast cells.
    Mol Cell Biol. 1992 Jul;12(7):3050-9 PMID: 1320195
  8. 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 Jan 25;266(3):1961-8 PMID: 1671046
  9. DNA polymerase epsilon: in search of a function.
    Trends Biochem Sci. 1992 Feb;17(2):55-8 PMID: 1566327
  10. Mechanism of elongation of primed DNA by DNA polymerase delta, proliferating cell nuclear antigen, and activator 1.
    Proc Natl Acad Sci U S A. 1990 Aug;87(15):5672-6 PMID: 1974050
  11. Purification of a cellular replication factor, RF-C, that is required for coordinated synthesis of leading and lagging strands during simian virus 40 DNA replication in vitro.
    Mol Cell Biol. 1989 Feb;9(2):609-19 PMID: 2565531
  12. Purification of a DNA polymerase-DNA primase complex from calf thymus glands.
    J Biol Chem. 1984 Dec 10;259(23):14679-87 PMID: 6094569
  13. Large-scale overproduction and rapid purification of the Escherichia coli ssb gene product. Expression of the ssb gene under lambda PL control.
    Biochemistry. 1986 Jan 14;25(1):21-5 PMID: 3006753
  14. Multiple replication factors augment DNA synthesis by the two eukaryotic DNA polymerases, alpha and delta.
    EMBO J. 1989 Dec 1;8(12):3883-9 PMID: 2573521
  15. Mammalian cyclin/PCNA (DNA polymerase delta auxiliary protein) stimulates processive DNA synthesis by yeast DNA polymerase III.
    Nucleic Acids Res. 1988 Jul 25;16(14A):6297-307 PMID: 2899883
  16. DNA unwinding activity of replication protein A.
    FEBS Lett. 1992 Aug 24;308(3):240-4 PMID: 1324186
  17. Replication factors required for SV40 DNA replication in vitro. I. DNA structure-specific recognition of a primer-template junction by eukaryotic DNA polymerases and their accessory proteins.
    J Biol Chem. 1991 Jan 25;266(3):1950-60 PMID: 1671045
  18. Interaction of DNA polymerase alpha-primase with cellular replication protein A and SV40 T antigen.
    EMBO J. 1992 Feb;11(2):769-76 PMID: 1311258
  19. Calf thymus RF-C as an essential component for DNA polymerase delta and epsilon holoenzymes function.
    Nucleic Acids Res. 1992 Aug 25;20(16):4159-65 PMID: 1354854
  20. cdc2 family kinases phosphorylate a human cell DNA replication factor, RPA, and activate DNA replication.
    EMBO J. 1992 Jun;11(6):2189-99 PMID: 1318195
  21. Multiple functions of human single-stranded-DNA binding protein in simian virus 40 DNA replication: single-strand stabilization and stimulation of DNA polymerases alpha and delta.
    Proc Natl Acad Sci U S A. 1989 Dec;86(24):9757-61 PMID: 2557626
  22. Synthesis of DNA by DNA polymerase epsilon in vitro.
    J Biol Chem. 1991 Nov 25;266(33):22707-17 PMID: 1682323
  23. Purification and characterization of replication protein A, a cellular protein required for in vitro replication of simian virus 40 DNA.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2523-7 PMID: 2833742
  24. Functional identity of proliferating cell nuclear antigen and a DNA polymerase-delta auxiliary protein.
    Nature. 1987 Apr 2-8;326(6112):517-20 PMID: 2882424
  25. Prokaryotic DNA replication mechanisms.
    Philos Trans R Soc Lond B Biol Sci. 1987 Dec 15;317(1187):395-420 PMID: 2894677
  26. Identification of replication factor C from Saccharomyces cerevisiae: a component of the leading-strand DNA replication complex.
    Mol Cell Biol. 1992 Jan;12(1):155-63 PMID: 1346062
  27. 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 Nov 25;266(33):22698-706 PMID: 1682322
  28. Immunoaffinity-purified DNA polymerase alpha displays novel properties.
    Biochemistry. 1987 Dec 15;26(25):8458-66 PMID: 3442669
  29. Eukaryotic DNA polymerases.
    Annu Rev Biochem. 1991;60:513-52 PMID: 1883203
  30. Sequence and expression in Escherichia coli of the 40-kDa subunit of activator 1 (replication factor C) of HeLa cells.
    Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2516-20 PMID: 1313560
  31. DNA Polymerase III holoenzyme of Escherichia coli. IV. The holoenzyme is an asymmetric dimer with twin active sites.
    J Biol Chem. 1988 May 15;263(14):6570-8 PMID: 3283128
  32. Revised nomenclature for eukaryotic DNA polymerases.
    Eur J Biochem. 1990 Aug 17;191(3):617-8 PMID: 2390988
  33. Biochemical and functional comparison of DNA polymerases alpha, delta, and epsilon from calf thymus.
    J Biol Chem. 1991 Jun 5;266(16):10420-8 PMID: 1674744
  34. Eukaryotic DNA replication. Enzymes and proteins acting at the fork.
    Eur J Biochem. 1990 Dec 27;194(3):699-712 PMID: 2269294
Article Info
Journal
Chromosoma
Abbr.
Chromosoma
ISSN
0009-5915
DOI
10.1007/BF02451797
Published
1992-00-00
Pages
S133-41
Language
English
Region
Austria
NLM ID
2985138R
Subset
IM
External Links
PubMed source DOI: 10.1007/BF02451797 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: [email protected]

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