Yeast open reading frame YCR14C encodes a DNA β-polymerase-like enzyme

Rajendra Prasad, Steven Widen, Rakesh K. Singhal, John Watkins, Louise Prakash, Samuel H. Wilson

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

We have shown by activity gel that overexpression in E.coli of a yeast chromosome 3 open reading frame (ORF) designated YCR14C and bearing homology to mammalian DNA polymerases βresults in a new DNA polymerase in the host cells. The molecular mass of this enzyme corresponded to the YCR14C-predicted 67 kDa protein, and NH2-terminal amino acid sequencing confirmed that the expressed protein was encoded by the yeast ORF. This new yeast DNA polymerase was purified to homogeneity from E.coli. In a fashion similar to that of mammalian β-polymerases, the purified yeast enzyme exhibited distributive DNA synthesis on DNA substrate with a single-stranded template and processive gap-filling synthesis on a short-gapped DNA substrate. Activity of this yeast β-polymerase-like enzyme was sensitive to the β-polymerase inhibitor ddNTP and resistant to both 1mM NEM and neutralizing antibody to E.coli DNA polymerase I. These results, therefore, indicate that YCR14C encodes a DNA β-polymerase-like enzyme in yeast, and we name it DNA polymerase IV. Yeast strains harboring a deletion mutation of the pol IV gene are viable, they exhibit no increase in sensitivity to ultraviolet light, ionizing radiation or alkylating agents, and sporulation and spore viability are not affected in the mutant.

Original languageEnglish (US)
Pages (from-to)5301-5307
Number of pages7
JournalNucleic Acids Research
Volume21
Issue number23
StatePublished - Nov 25 1993

Fingerprint

DNA-Directed DNA Polymerase
Yeast
Open Reading Frames
Enzymes
DNA
Yeasts
Escherichia coli
Escherichia Coli
Bearings (structural)
Dideoxynucleotides
Substrate
pol Genes
Synthesis
DNA Polymerase beta
Protein
DNA Polymerase I
Chromosomes, Human, Pair 3
Sequence Deletion
Alkylating Agents
Ionizing radiation

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Prasad, R., Widen, S., Singhal, R. K., Watkins, J., Prakash, L., & Wilson, S. H. (1993). Yeast open reading frame YCR14C encodes a DNA β-polymerase-like enzyme. Nucleic Acids Research, 21(23), 5301-5307.

Yeast open reading frame YCR14C encodes a DNA β-polymerase-like enzyme. / Prasad, Rajendra; Widen, Steven; Singhal, Rakesh K.; Watkins, John; Prakash, Louise; Wilson, Samuel H.

In: Nucleic Acids Research, Vol. 21, No. 23, 25.11.1993, p. 5301-5307.

Research output: Contribution to journalArticle

Prasad, R, Widen, S, Singhal, RK, Watkins, J, Prakash, L & Wilson, SH 1993, 'Yeast open reading frame YCR14C encodes a DNA β-polymerase-like enzyme', Nucleic Acids Research, vol. 21, no. 23, pp. 5301-5307.
Prasad R, Widen S, Singhal RK, Watkins J, Prakash L, Wilson SH. Yeast open reading frame YCR14C encodes a DNA β-polymerase-like enzyme. Nucleic Acids Research. 1993 Nov 25;21(23):5301-5307.
Prasad, Rajendra ; Widen, Steven ; Singhal, Rakesh K. ; Watkins, John ; Prakash, Louise ; Wilson, Samuel H. / Yeast open reading frame YCR14C encodes a DNA β-polymerase-like enzyme. In: Nucleic Acids Research. 1993 ; Vol. 21, No. 23. pp. 5301-5307.
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