TY - JOUR
T1 - Molecular Analyses of an Unusual Translesion DNA Polymerase from Methanosarcina acetivorans C2A
AU - Lin, Li Jung
AU - Yoshinaga, Aya
AU - Lin, Yuyen
AU - Guzman, Claudia
AU - Chen, Yi Hsing
AU - Mei, Shou
AU - Lagunas, Angelica M.
AU - Koike, Satoshi
AU - Iwai, Shigenori
AU - Spies, M. Ashley
AU - Nair, Satish K.
AU - Mackie, Roderick I.
AU - Ishino, Yoshizumi
AU - Cann, Isaac K.O.
N1 - Funding Information:
This research was supported by the National Science Foundation through grant MCB 0238451 (to I.K.O.C.). We thank Dr. William Metcalf, University of Illinois at Urbana-Champaign, for providing M. acetivorans genomic DNA.
PY - 2010/3/19
Y1 - 2010/3/19
N2 - 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.
AB - 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.
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U2 - 10.1016/j.jmb.2010.01.007
DO - 10.1016/j.jmb.2010.01.007
M3 - Article
C2 - 20080107
AN - SCOPUS:77249113717
SN - 0022-2836
VL - 397
SP - 13
EP - 30
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 1
ER -