Identification of a higher molecular weight DNA polymerase α catalytic polypeptide in monkey cells by monoclonal antibody

E. Karawya, J. Swack, W. Albert, J. Fedorko, J. D. Minna, S. H. Wilson

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

A monoclonal antibody against purified calf DNA polymerase α (deoxynucleosidetriphosphate:DNA deoxy-nucleotidyltransferase, EC 2.7.7.7) was used to immunoprecipitate proteins from a crude soluble extract of growing monkey BSC-1 cells. Immunoprecipitates contained familiar DNA polymerase α catalytic polypeptides of M(r)S ≃ 115,000 and 70,000 and also a M(r) 40,000 catalytic polypeptide; the major component in the immunoprecipitates, however, was a polypeptide M(r) ≃ 190,000 not previously identified as a DNA polymerase. This protein was capable of DNA polymerase activity after electroelution from NaDodSO4/polyacrylamide gels and renaturation. The highly purified enzyme so obtained was active with poly(dT)·oligo(rA) as template·primer, resistant to dideoxy TTP (ddTTP), and inhibited by aphidicolin and butylphenyldeoxyguanosine 5'-triphosphate, thus identifying it as a DNA polymerase α. The results indicate that a polypeptide of M(r) ≃ 190,000 is an abundant component among DNA polymerase α catalytic polypeptides in growing monkey cells.

Original languageEnglish (US)
Pages (from-to)7777-7781
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume81
Issue number24 I
DOIs
StatePublished - 1984

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DNA-Directed DNA Polymerase
Haplorhini
Molecular Weight
Monoclonal Antibodies
Peptides
DNA Nucleotidyltransferases
Aphidicolin
Complex Mixtures
Proteins
Enzymes

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Identification of a higher molecular weight DNA polymerase α catalytic polypeptide in monkey cells by monoclonal antibody. / Karawya, E.; Swack, J.; Albert, W.; Fedorko, J.; Minna, J. D.; Wilson, S. H.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 81, No. 24 I, 1984, p. 7777-7781.

Research output: Contribution to journalArticle

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AU - Karawya, E.

AU - Swack, J.

AU - Albert, W.

AU - Fedorko, J.

AU - Minna, J. D.

AU - Wilson, S. H.

PY - 1984

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N2 - A monoclonal antibody against purified calf DNA polymerase α (deoxynucleosidetriphosphate:DNA deoxy-nucleotidyltransferase, EC 2.7.7.7) was used to immunoprecipitate proteins from a crude soluble extract of growing monkey BSC-1 cells. Immunoprecipitates contained familiar DNA polymerase α catalytic polypeptides of M(r)S ≃ 115,000 and 70,000 and also a M(r) 40,000 catalytic polypeptide; the major component in the immunoprecipitates, however, was a polypeptide M(r) ≃ 190,000 not previously identified as a DNA polymerase. This protein was capable of DNA polymerase activity after electroelution from NaDodSO4/polyacrylamide gels and renaturation. The highly purified enzyme so obtained was active with poly(dT)·oligo(rA) as template·primer, resistant to dideoxy TTP (ddTTP), and inhibited by aphidicolin and butylphenyldeoxyguanosine 5'-triphosphate, thus identifying it as a DNA polymerase α. The results indicate that a polypeptide of M(r) ≃ 190,000 is an abundant component among DNA polymerase α catalytic polypeptides in growing monkey cells.

AB - A monoclonal antibody against purified calf DNA polymerase α (deoxynucleosidetriphosphate:DNA deoxy-nucleotidyltransferase, EC 2.7.7.7) was used to immunoprecipitate proteins from a crude soluble extract of growing monkey BSC-1 cells. Immunoprecipitates contained familiar DNA polymerase α catalytic polypeptides of M(r)S ≃ 115,000 and 70,000 and also a M(r) 40,000 catalytic polypeptide; the major component in the immunoprecipitates, however, was a polypeptide M(r) ≃ 190,000 not previously identified as a DNA polymerase. This protein was capable of DNA polymerase activity after electroelution from NaDodSO4/polyacrylamide gels and renaturation. The highly purified enzyme so obtained was active with poly(dT)·oligo(rA) as template·primer, resistant to dideoxy TTP (ddTTP), and inhibited by aphidicolin and butylphenyldeoxyguanosine 5'-triphosphate, thus identifying it as a DNA polymerase α. The results indicate that a polypeptide of M(r) ≃ 190,000 is an abundant component among DNA polymerase α catalytic polypeptides in growing monkey cells.

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