Characterization of an ATP-dependent DNA ligase encoded by Chlorella virus PBCV-1 Journal Article
| Authors: | Ho, C. K.; Van Etten, J. L.; Shuman, S. |
| Article Title: | Characterization of an ATP-dependent DNA ligase encoded by Chlorella virus PBCV-1 |
| 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. |
| Keywords: | nonhuman; animals; gene expression; dna; amino acid sequence; molecular sequence data; sequence homology, amino acid; kinetics; recombinant fusion proteins; escherichia coli; substrate specificity; recombinant protein; adenosine triphosphate; polydeoxyribonucleotide synthase; enzyme specificity; enzyme substrate complex; guanosine triphosphate; virus genome; dna nick translation; dna ligases; viral proteins; nucleotidyltransferase; cations, divalent; magnesium; rna capping; nucleoside triphosphate; adenosine monophosphate; dna virus; chlorella; humans; priority journal; article; phycodnaviridae |
| Journal Title: | Journal of Virology |
| Volume: | 71 |
| Issue: | 3 |
| ISSN: | 0022-538X |
| Publisher: | American Society for Microbiology |
| Date Published: | 1997-03-01 |
| Start Page: | 1931 |
| End Page: | 1937 |
| Language: | English |
| PUBMED: | 9032324 |
| PROVIDER: | scopus |
| PMCID: | PMC191272 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 17 March 2017 -- Source: Scopus |
