Characterization of an ATP-dependent DNA ligase encoded by Chlorella virus PBCV-1

C. Kiong Ho, James L. Van Etten, Stewart Shuman

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

We report that Chlorella virus PBCV-1 encodes a 298-amino-acid ATP- dependent DNA ligase. The PBCV-1 enzyme is the smallest member of the covalent nucleotidyl transferase superfamily, which includes the ATP- dependent polynucleotide ligases and the GTP-dependent RNA capping enzymes. The specificity of PBCV-1 DNA ligase was investigated by using purified recombinant protein. The enzyme catalyzed efficient strand joining on a singly nicked DNA in the presence of magnesium and ATP (K(m), 75 μM). Other nucleoside triphosphates or deoxynucleoside triphosphates could not substitute for ATP. PBCV-1 ligase was unable to ligate across a 2-nucleotide gap and ligated poorly across a 1-nucleotide gap. A native gel mobility shift assay showed that PBCV-1 DNA ligase discriminated between nicked and gapped DNAs at the substrate-binding step. These findings underscore the importance of a properly positioned 3' OH acceptor terminus in substrate recognition and reaction chemistry.

Original languageEnglish (US)
Pages (from-to)1931-1937
Number of pages7
JournalJournal of virology
Volume71
Issue number3
StatePublished - Mar 1 1997

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Paramecium bursaria Chlorella virus 1
Chlorella
ligases
DNA Ligases
Adenosine Triphosphate
Viruses
viruses
Polynucleotide Ligases
DNA
Enzymes
Nucleotides
Electrophoretic Mobility Shift Assay
Ligases
Transferases
Guanosine Triphosphate
Nucleosides
reaction chemistry
Recombinant Proteins
Magnesium
enzymes

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Characterization of an ATP-dependent DNA ligase encoded by Chlorella virus PBCV-1. / Ho, C. Kiong; Van Etten, James L.; Shuman, Stewart.

In: Journal of virology, Vol. 71, No. 3, 01.03.1997, p. 1931-1937.

Research output: Contribution to journalArticle

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