Reference: Plevani P, et al. (1985) Polypeptide structure of DNA primase from a yeast DNA polymerase-primase complex. J Biol Chem 260(11):7102-7

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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.

Reference Type
Journal Article | Research Support, Non-U.S. Gov't | Research Support, U.S. Gov't, Non-P.H.S. | Research Support, U.S. Gov't, P.H.S.
Authors
Plevani P, Foiani M, Valsasnini P, Badaracco G, Cheriathundam E, Chang LM
Primary Lit For
PRI1 | POL12 | PRI2 | POL1 | DNA polymerase alpha:primase complex

Gene Ontology Annotations


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Gene/Complex Qualifier Gene Ontology Term Aspect Annotation Extension Evidence Method Source Assigned On Reference

Interaction Annotations


Genetic Interactions

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Interactor Interactor Allele Assay Annotation Action Phenotype SGA score P-value Source Reference

Physical Interactions 3 entries for 4 genes

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InteractorInteractorAssayAnnotationActionModification
PRI1POL1Co-purificationmanually curatedBait-HitNo Modification
PRI1POL2Co-purificationmanually curatedBait-HitNo Modification
PRI1PRI2Co-purificationmanually curatedBait-HitNo Modification
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