T4 endonuclease V promotes the formation of multimeric DNA structures

Robert (Stephen) Lloyd, M. L. Dodson, E. A. Gruskin, D. L. Robberson

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Electron microscopy of UV-irradiated circular DNA molecules which had been treated with T4 endonuclease V revealed the formation of multimeric DNA structures in addition to the expected conversion of the superhelical DNA molecules into nicked circular and linear forms. The multimeric DNA molecules could be distinguished in electron micrographs from catenated molecules which were present in the original DNA preparation by a combination of rotary and single angle heavy metal shadowing. The complexity and frequency of these structures increased with time of reaction with endonuclease V. Their formation, as well as the endonuclease activity of enzyme, was dependent on UV irradiation of the DNA, and the complexes could be disrupted by prior phenol extraction and ethanol precipitation. Preparations of endonuclease V estimated to be 98% pure by mass promoted the same complex formation between DNA molecule as did preparations estimated to be only 5-10% pure. In addition to these intermolecular structures, the formation of complexes between regions on the same DNA molecules was manifest as discrete double-stranded 'loops' 200-300 base pairs in length. DNA 'bubble structures' were also observed and may represent folding of the 'loops' onto adjacent segments of DNA. These results suggest that at least one active form of T4 endonuclease V may be a multimetric complex of enzyme molecules in association with DNA.

Original languageEnglish (US)
Pages (from-to)109-115
Number of pages7
JournalMutation Research DNA Repair Reports
Volume183
Issue number2
DOIs
StatePublished - 1987
Externally publishedYes

Fingerprint

DNA
Deoxyribonuclease (Pyrimidine Dimer)
Superhelical DNA
phage T4 endonuclease V
Circular DNA
Endonucleases
Enzymes
Heavy Metals
Phenol
Base Pairing
Electron Microscopy
Ethanol
Electrons

Keywords

  • Bacteriophage T4
  • DNA
  • Endonuclease V
  • multimetric structures
  • Pyrimidine dimers
  • T4
  • T4 endonuclease V
  • UV-induced

ASJC Scopus subject areas

  • Medicine(all)

Cite this

T4 endonuclease V promotes the formation of multimeric DNA structures. / Lloyd, Robert (Stephen); Dodson, M. L.; Gruskin, E. A.; Robberson, D. L.

In: Mutation Research DNA Repair Reports, Vol. 183, No. 2, 1987, p. 109-115.

Research output: Contribution to journalArticle

Lloyd, Robert (Stephen) ; Dodson, M. L. ; Gruskin, E. A. ; Robberson, D. L. / T4 endonuclease V promotes the formation of multimeric DNA structures. In: Mutation Research DNA Repair Reports. 1987 ; Vol. 183, No. 2. pp. 109-115.
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