Molecular analyses of an unusual translesion DNA polymerase from Methanosarcina acetivorans C2A

J Mol Biol. 2010 Mar 19;397(1):13-30. doi: 10.1016/j.jmb.2010.01.007. Epub 2010 Jan 18.


The domain Archaea is composed of several subdomains, and prominent among them are the Crenarchaeota and the Euryarchaeota. Biochemically characterized archaeal family Y DNA polymerases (Pols) or DinB homologs, to date, are all from crenarchaeal organisms, especially the genus Sulfolobus. Here, we demonstrate that archaeal family Y Pols fall into five clusters based on phylogenetic analysis. MacDinB-1, the homolog from the euryarchaeon Methanosarcina acetivorans that is characterized in this study, belongs to cluster II. Therefore, MacDinB-1 is different from the Sulfolobus DinB proteins, which are members of cluster I. In addition to translesion DNA synthesis activity, MacDinB-1 synthesized unusually long products ( approximately 7.2 kb) in the presence of its cognate proliferating cell nuclear antigen (PCNA). The PCNA-interacting site in MacDinB-1 was identified by mutational analysis in a C-terminally located heptapeptide akin to a PIP (PCNA-interacting protein) box. In vitro assays from the present report suggested that MacDinB-1 works in an error-free mode to repair cyclobutane pyrimidine dimers. This study on a euryarchaeal DinB homolog provides important insights into the functional diversity of the family Y Pols, and the availability of a genetic system for this archaeon should allow subsequent elucidation of the physiological significance of this enzyme in M. acetivorans cells.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adenine / metabolism
  • Amino Acid Motifs
  • Amino Acid Sequence
  • Archaeal Proteins / chemistry
  • Archaeal Proteins / isolation & purification
  • DNA Damage
  • DNA Primers / metabolism
  • DNA, Archaeal / biosynthesis
  • DNA-Directed DNA Polymerase / isolation & purification
  • DNA-Directed DNA Polymerase / metabolism*
  • Humans
  • Methanosarcina / enzymology*
  • Molecular Sequence Data
  • Multigene Family / genetics
  • Mutation / genetics
  • Phylogeny
  • Proliferating Cell Nuclear Antigen / metabolism
  • Protein Binding
  • Sequence Alignment
  • Sequence Homology, Amino Acid


  • Archaeal Proteins
  • DNA Primers
  • DNA, Archaeal
  • Proliferating Cell Nuclear Antigen
  • DNA-Directed DNA Polymerase
  • Adenine