This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ho, C. K. Articles by Shuman, S. Search for Related Content PubMed PubMed Citation Articles by Ho, C. K. Articles by Shuman, S.

 Previous Article  |  Next Article 

J. Virol., 03 1997, 1931-1937, Vol 71, No. 3
Copyright © 1997, American Society for Microbiology

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

CK Ho, JL Van Etten and S Shuman
Molecular Biology Program, Sloan-Kettering Institute, New York, New York 10021, USA.

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 (Km, 75 microM). 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.

This article has been cited by other articles:

• Brooks, M. A., Meslet-Cladiere, L., Graille, M., Kuhn, J., Blondeau, K., Myllykallio, H., van Tilbeurgh, H. (2008). The structure of an archaeal homodimeric ligase which has RNA circularization activity. Protein Sci. 17: 1336-1345 [Abstract] [Full Text]  
• Kiyonari, S., Takayama, K., Nishida, H., Ishino, Y. (2006). Identification of a Novel Binding Motif in Pyrococcus furiosus DNA Ligase for the Functional Interaction with Proliferating Cell Nuclear Antigen. J. Biol. Chem. 281: 28023-28032 [Abstract] [Full Text]  
• Nandakumar, J., Ho, C. K., Lima, C. D., Shuman, S. (2004). RNA Substrate Specificity and Structure-guided Mutational Analysis of Bacteriophage T4 RNA Ligase 2. J. Biol. Chem. 279: 31337-31347 [Abstract] [Full Text]  
• Odell, M., Malinina, L., Sriskanda, V., Teplova, M., Shuman, S. (2003). Analysis of the DNA joining repertoire of Chlorella virus DNA ligase and a new crystal structure of the ligase-adenylate intermediate. Nucleic Acids Res 31: 5090-5100 [Abstract] [Full Text]  
• Sriskanda, V., Shuman, S. (2002). Role of Nucleotidyl Transferase Motif V in Strand Joining by Chlorella Virus DNA Ligase. J. Biol. Chem. 277: 9661-9667 [Abstract] [Full Text]  
• Sriskanda, V., Shuman, S. (2002). Role of nucleotidyltransferase motifs I, III and IV in the catalysis of phosphodiester bond formation by Chlorella virus DNA ligase. Nucleic Acids Res 30: 903-911 [Abstract] [Full Text]  
• Sriskanda, V., Kelman, Z., Hurwitz, J., Shuman, S. (2000). Characterization of an ATP-dependent DNA ligase from the thermophilic archaeon Methanobacterium thermoautotrophicum. Nucleic Acids Res 28: 2221-2228 [Abstract] [Full Text]  
• Tong, J., Barany, F., Cao, W. (2000). Ligation reaction specificities of an NAD+-dependent DNA ligase from the hyperthermophile Aquifex aeolicus. Nucleic Acids Res 28: 1447-1454 [Abstract] [Full Text]  
• Lavrukhin, O. V., Fortune, J. M., Wood, T. G., Burbank, D. E., Van Etten, J. L., Osheroff, N., Lloyd, R. S. (2000). Topoisomerase II from Chlorella Virus PBCV-1. CHARACTERIZATION OF THE SMALLEST KNOWN TYPE II TOPOISOMERASE. J. Biol. Chem. 275: 6915-6921 [Abstract] [Full Text]  
• Ciarrocchi, G., MacPhee, D. G., Deady, L. W., Tilley, L. (1999). Specific Inhibition of the Eubacterial DNA Ligase by Arylamino Compounds. Antimicrob. Agents Chemother. 43: 2766-2772 [Abstract] [Full Text]  
• Odell, M., Shuman, S. (1999). Footprinting of Chlorella Virus DNA Ligase Bound at a Nick in Duplex DNA. J. Biol. Chem. 274: 14032-14039 [Abstract] [Full Text]  
• Pearson, M. N., Rohrmann, G. F. (1998). Characterization of a Baculovirus-Encoded ATP-Dependent DNA Ligase. J. Virol. 72: 9142-9149 [Abstract] [Full Text]