title: Molecular analyses of an unusual translesion DNA polymerase from Methanosarcina acetivorans C2A.
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 ( 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.
authors.affiliation: Department of Animal Sciences, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
authors.forename: Li-Jung
authors.initials: LJ
authors.lastname: Lin
authors.forename: Aya
authors.initials: A
authors.lastname: Yoshinaga
authors.forename: Yuyen
authors.initials: Y
authors.lastname: Lin
authors.forename: Claudia
authors.initials: C
authors.lastname: Guzman
authors.forename: Yi-Hsing
authors.initials: YH
authors.lastname: Chen
authors.forename: Shou
authors.initials: S
authors.lastname: Mei
authors.forename: Angelica M
authors.initials: AM
authors.lastname: Lagunas
authors.forename: Satoshi
authors.initials: S
authors.lastname: Koike
authors.forename: Shigenori
authors.initials: S
authors.lastname: Iwai
authors.forename: M Ashley
authors.initials: MA
authors.lastname: Spies
authors.forename: Satish K
authors.initials: SK
authors.lastname: Nair
authors.forename: Roderick I
authors.initials: RI
authors.lastname: Mackie
authors.forename: Yoshizumi
authors.initials: Y
authors.lastname: Ishino
authors.forename: Isaac K O
authors.initials: IK
authors.lastname: Cann
mesh_terms: D000225:Adenine / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry / Q000302:isolation & purification; D004249:DNA Damage; D017931:DNA Primers / Q000378:metabolism; D019641:DNA, Archaeal / Q000096:biosynthesis; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D017020:Methanosarcina / Q000201:enzymology*; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics; D009154:Mutation / Q000235:genetics; D010802:Phylogeny; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D011485:Protein Binding; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D019843:Archaeal Proteins; D017931:DNA Primers; D019641:DNA, Archaeal; D018809:Proliferating Cell Nuclear Antigen; D004259:DNA-Directed DNA Polymerase; D000225:Adenine