Literature DB >> 3888995

Polypeptide structure of DNA primase from a yeast DNA polymerase-primase complex.

P Plevani, M Foiani, P Valsasnini, G Badaracco, E Cheriathundam, L M Chang.   

Abstract

An immunoaffinity chromatographic procedure was developed to purify DNA polymerase-DNA primase complex from crude soluble extracts of yeast cells. The immunoabsorbent column is made of mouse monoclonal antibody to yeast DNA polymerase I covalently linked to Protein A-Sepharose. Purification of the complex involves binding of the complex to the immunoabsorbent column and elution with concentrated MgCl2 solutions. After rebinding to the monoclonal antibody column free primase activity is selectively eluted with a lower concentration of MgCl2. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate showed the presence of five major peptides, p180, p140, p74, p58, and p48 in the immunoaffinity-purified DNA polymerase-DNA primase complex. Free primase and free polymerase fractions obtained by fractionation on the immunoabsorbent column were analyzed on activity gels and immunoblots. These analyses showed that p180 and p140 are DNA polymerase peptides. Two polypeptides of 58 and 48 kDa co-fractionated with the free yeast DNA primase. From sucrose gradient analysis we estimate a molecular weight of 110 kDa for the native DNA primase.

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Year:  1985        PMID： 3888995

Source DB:  PubMed          Journal:  J Biol Chem        ISSN： 0021-9258            Impact factor:   5.157


  26 in total

1.  Pea chloroplast DNA primase: characterization and role in initiation of replication.

Authors:  B L Nielsen; V K Rajasekhar; K K Tewari
Journal:  Plant Mol Biol       Date:  1991-06       Impact factor: 4.076

2.  Cyclin B-cdk1 kinase stimulates ORC- and Cdc6-independent steps of semiconservative plasmid replication in yeast nuclear extracts.

Authors:  B P Duncker; P Pasero; D Braguglia; P Heun; M Weinreich; S M Gasser
Journal:  Mol Cell Biol       Date:  1999-02       Impact factor: 4.272

3.  Fidelity of DNA polymerase I and the DNA polymerase I-DNA primase complex from Saccharomyces cerevisiae.

Authors:  T A Kunkel; R K Hamatake; J Motto-Fox; M P Fitzgerald; A Sugino
Journal:  Mol Cell Biol       Date:  1989-10       Impact factor: 4.272

4.  Identification of the yeast DNA polymerase I gene with antibody probes.

Authors:  G Lucchini; A Brandazza; G Badaracco; M Bianchi; P Plevani
Journal:  Curr Genet       Date:  1985       Impact factor: 3.886

Review 5.  Yeast chromosome replication and segregation.

Authors:  C S Newlon
Journal:  Microbiol Rev       Date:  1988-12

6.  Resolution and purification of free primase activity from the DNA primase-polymerase alpha complex of HeLa cells.

Authors:  J K Vishwanatha; E F Baril
Journal:  Nucleic Acids Res       Date:  1986-11-11       Impact factor: 16.971

7.  DNA polymerase I gene of Saccharomyces cerevisiae: nucleotide sequence, mapping of a temperature-sensitive mutation, and protein homology with other DNA polymerases.

Authors:  A Pizzagalli; P Valsasnini; P Plevani; G Lucchini
Journal:  Proc Natl Acad Sci U S A       Date:  1988-06       Impact factor: 11.205

8.  Cell cycle-dependent phosphorylation and dephosphorylation of the yeast DNA polymerase alpha-primase B subunit.

Authors:  M Foiani; G Liberi; G Lucchini; P Plevani
Journal:  Mol Cell Biol       Date:  1995-02       Impact factor: 4.272

9.  Stimulation of human neuroblastoma DNA polymerase alpha and primase activities by a protein factor isolated from rat liver chromatin.

Authors:  S Takada; A Torres-Rosado; S Ray; S Basu
Journal:  Proc Natl Acad Sci U S A       Date:  1986-12       Impact factor: 11.205

10.  Conditional mutations in the yeast DNA primase genes affect different aspects of DNA metabolism and interactions in the DNA polymerase alpha-primase complex.

Authors:  M P Longhese; L Jovine; P Plevani; G Lucchini
Journal:  Genetics       Date:  1993-02       Impact factor: 4.562
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