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Microbiology and Immunology

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A DNA polymerase from Ustilago maydis. 1. Purification and properties of the polymerase activity.

TitleA DNA polymerase from Ustilago maydis. 1. Purification and properties of the polymerase activity.
Publication TypeJournal Article
Year of Publication1976
AuthorsBanks GR, Holloman WK, Kairis MV, Spanos A, Yarranton GT
JournalEur J Biochem
Volume62
Issue1
Pagination131-42
Date Published1976 Feb 2
ISSN0014-2956
KeywordsBasidiomycota, Cations, Divalent, Cations, Monovalent, DNA Nucleotidyltransferases, DNA Replication, Kinetics, Macromolecular Substances, Molecular Weight, Templates, Genetic, Ustilago
Abstract

A DNA polymerase from Ustilago maydis has been purified to apparent homogeneity. The native enzyme possesses a subunit structure consisting of 50000 and 55000-dalton monomers. The apparent sedimentation coefficient of the polymerase activity in the absence of salt is 8.4 S (Mr=180000-200000), that in its presence (0.6 M NaCl or 0.12 M KCl) being 6.3 S (Mr=80000-100000). Low concentrations of EDTA also converted the 8.4-S to a 6.3-S form, whereas magnesium ions catalysed the reverse association. The enzyme has an absolute requirement for both a DNA or RNA template and a DNA primer. For homopolymer templates the primer requirement was satisified by a short complementary oligodeoxynucleotide, but oligoribonucleotides were extremely inefficient primers. With the template-primer poly(dA) X (dT)12, the enzyme added an average of 50 dTMP nucleotides on to each primer molecule, whereas with poly(rA) X (dT)12, this figure was 300. The enzyme also possesses an associated deoxyribonuclease activity. No other DNA polymerase activity was detected in cell-free extracts of U. maydis.

Alternate JournalEur. J. Biochem.
PubMed ID1248475

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