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

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

来源：Journal of Molecular Biology, 2010, 397(1): 13-30.

DOI：10.1016/j.jmb.2010.01.007

• translesion DNA synthesis
• DinB
• Archaea
• family Y polymerase
• PCNA

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 (similar to 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. hi 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.

• 出版日期2010-3-19