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DNA polymerase lambda directly binds to proliferating cell nuclear antigen through its confined C‐terminal region
Author(s) -
Shimazaki Noriko,
Yazaki Takaya,
Kubota Takashi,
Sato Asami,
Nakamura Ayako,
Kurei Shunsuke,
Toji Shingo,
Tamai Katsuyuki,
Koiwai Osamu
Publication year - 2005
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1111/j.1365-2443.2005.00868.x
Subject(s) - proliferating cell nuclear antigen , dna polymerase , biology , dna polymerase delta , processivity , microbiology and biotechnology , dna replication , polymerase , dna clamp , dna polymerase ii , dna , biochemistry , gene , reverse transcriptase , rna
DNA polymerase lambda (Pol λ) was recently identified as a new member of the family X of DNA polymerases. Here, we show that Pol λ directly binds to proliferating cell nuclear antigen (PCNA), an auxiliary protein for DNA replication and repair enzymes, both in vitro and in vivo . A pull‐down assay using deletion mutants of Pol λ showed that the confined C‐terminal region of Pol λ directly binds to PCNA. Furthermore, a synthetic peptide of 20‐mers derived from the C‐terminal region of Pol λ competes with full‐length Pol λ for binding to PCNA. The residues between amino acids 518 and 537 of Pol λ are required for binding to PCNA, and are different from the consensus PCNA interacting motif (PIM). Pol λ associates with PCNA in vivo by immunoprecipitation analysis and EGFP‐tagged Pol λ co‐localizes with PCNA as spots within a nucleus using fluorescent microscopy. Through direct binding, PCNA suppressed the distributive nucleotidyltransferase activity of Pol λ. Pol µ, which also belongs to the family X of DNA polymerases, binds to PCNA by a pivotal amino acid residue.