Molecular Analyses of an Unusual Translesion DNA Polymerase from Methanosarcina acetivorans C2A

Li Jung Lin, Aya Yoshinaga, Yuyen Lin, Claudia Guzman, Yi Hsing Chen, Shou Mei, Angelica M. Lagunas, Satoshi Koike, Shigenori Iwai, M. Ashley Spies, Satish K. Nair, Roderick I. Mackie, Yoshizumi Ishino, Isaac K.O. Cann

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Abstract

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 (∼ 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.

Original languageEnglish
Pages (from-to)13-30
Number of pages18
JournalJournal of Molecular Biology
Volume397
Issue number1
DOIs
Publication statusPublished - Mar 19 2010

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All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Cite this

Lin, L. J., Yoshinaga, A., Lin, Y., Guzman, C., Chen, Y. H., Mei, S., ... Cann, I. K. O. (2010). Molecular Analyses of an Unusual Translesion DNA Polymerase from Methanosarcina acetivorans C2A. Journal of Molecular Biology, 397(1), 13-30. https://doi.org/10.1016/j.jmb.2010.01.007