Immunoaffinity purification and properties of Drosophila melanogaster DNA polymerase alpha-primase complex.

Immunoaffinity purification and properties of Drosophila melanogaster DNA polymerase alpha-primase complex. Research Abstract Details 

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Immunoaffinity purification and properties of Drosophila melanogaster DNA polymerase alpha-primase complex. Abstract Text:

K Kuroda,R Kagiyama-Takahashi,T Shinomiya,

Two hybrid cell lines (DM88-5E12 and DM88-4C9) secreting monoclonal antibodies against DNA polymerase alpha-primase complex from Drosophila melanogaster Kc cells were established by immunizing mice with the complex partially purified by a conventional method. The IgG subclasses of both antibodies were IgG1. Both antibodies immunoprecipitated the DNA polymerase alpha-primase complex from D. melanogaster Kc cells. The DNA-polymerizing activity was neutralized by 4C9 antibody, but not by 5E12 antibody. The DNA priming activity was not neutralized by either antibody. These antibodies did not cross-react to HeLa DNA polymerase alpha-primase complex. A rapid, two-step purification of DNA polymerase alpha-primase complex from D. melanogaster Kc cell was carried out by 5E12 antibody column chromatography followed by single-stranded DNA cellulose column chromatography. The immunoaffinity-purified enzyme had both DNA-polymerizing and DNA-priming activities with the specific activities of 50,000 and 2,000 units/mg, respectively. The effects of aphidicolin, NEM, ddTTP, BuPdGTP, and DMSO on the enzyme activity showed that the purified enzyme was DNA polymerase alpha, but not DNA polymerase beta, gamma, or delta. The purified enzyme consisted of polypeptides with apparent molecular weights of 180 (and 145, 140, 130 kDa), 72, 63, 51, and 49 kDa. The 5E12 antibody was shown to bind to all the high-molecular-weight polypeptides, 180, 145, 140, and 130 kDa, by immuno-Western blotting analysis.

Immunoaffinity purification and properties of Drosophila melanogaster DNA polymerase alpha-primase complex. Publishing Authors By Initials

K Kuroda,R Kagiyama-Takahashi,T Shinomiya,

For similar enzymes and coenzymes: enzymes: transferases: phosphotransferases: nucleotidyltransferases: rna nucleotidyltransferases research abstracts see: enzymes and coenzymes: enzymes: transferases: phosphotransferases: nucleotidyltransferases: rna nucleotidyltransferases research

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Immunoaffinity purification and properties of Drosophila melanogaster DNA polymerase alpha-primase complex. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Journal of biochemistry

VOLUME: 108

Page Numbers: 926-33

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Dec

YEAR: 1990

Immunoaffinity purification and properties of Drosophila melanogaster DNA polymerase alpha-primase complex. Information

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LANGUAGE: eng

NlmUniqueID: 376600

Immunoaffinity purification and properties of Drosophila melanogaster DNA polymerase alpha-primase complex. Keywords Mesh Terms:

KEYWORDS: RNA Nucleotidyltransferases

MESH TERMS: isolation & purification

Chemical & Substance for Abstract: Immunoaffinity purification and properties of Drosophila melanogaster DNA polymerase alpha-primase complex. Information

Substance Name: RNA Nucleotidyltransferases

Registry Number: EC 2.7.7.-

Grant and Affiliation Information for Immunoaffinity purification and properties of Drosophila melanogaster DNA polymerase alpha-primase complex.

AFFILIATION: Mitsubishi Kasei Institute of Life Sciences, Tokyo.

Country: JAPAN

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MEDLINETA: J Biochem

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