DNA substrate specificity of DNA helicase E from calf thymus

John Turchi, Richard S. Murante, Robert A. Bambara

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

DNA helicase E from calf thymus has been characterized with respect to DNA substrate specificity. The helicase was capable of displacing DNA fragments up to 140 nucleotides in length, but was unable to displace a DNA fragment 322 nucleotides in length. DNA competition experiments revealed that helicase E was moderately processive for translocation on single strand M13mp18 DNA, and that the helicase would dissociate and rebind during a 15 minute reaction. Comparison of the rate of ATPase activity catalyzed by helicase E on single strand DNA substrates of different lengths, suggested a processivity consistent with the competition experiments. The helicase displayed a preference for displacing primers whose 5′ terminus was fully annealed as opposed to primers with a 12 nucleotide 5′ unannealed tail. The presence of a 12 nucleotide 3′ tail had no effect on the rate of displacement. DNA helicase E was capable of displacing a primer downstream of either a four nucleotide gap, a one nucleotide gap or a nick in the DNA substrate. Helicase E was inactive on a fully duplex DNA 30 base pairs in length. Calf thymus RP-A stimulated the DNA displacement activity of helicase E. These properties are consistent with a role for DNA helicase E in chromosomal DNA repair.

Original languageEnglish (US)
Pages (from-to)6075-6080
Number of pages6
JournalNucleic Acids Research
Volume20
Issue number22
StatePublished - Nov 25 1992
Externally publishedYes

Fingerprint

DNA Helicases
Thymus
Substrate Specificity
Thymus Gland
Specificity
DNA
Substrate
Nucleotides
Substrates
Tail
Fragment
ATP Synthase
Translocation
Repair
Experiment
Single-Stranded DNA Breaks
Calves
Base Pairing
DNA Repair
Adenosine Triphosphatases

ASJC Scopus subject areas

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

Cite this

Turchi, J., Murante, R. S., & Bambara, R. A. (1992). DNA substrate specificity of DNA helicase E from calf thymus. Nucleic Acids Research, 20(22), 6075-6080.

DNA substrate specificity of DNA helicase E from calf thymus. / Turchi, John; Murante, Richard S.; Bambara, Robert A.

In: Nucleic Acids Research, Vol. 20, No. 22, 25.11.1992, p. 6075-6080.

Research output: Contribution to journalArticle

Turchi, J, Murante, RS & Bambara, RA 1992, 'DNA substrate specificity of DNA helicase E from calf thymus', Nucleic Acids Research, vol. 20, no. 22, pp. 6075-6080.
Turchi, John ; Murante, Richard S. ; Bambara, Robert A. / DNA substrate specificity of DNA helicase E from calf thymus. In: Nucleic Acids Research. 1992 ; Vol. 20, No. 22. pp. 6075-6080.
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