title: The DNA binding domain of human DNA ligase I interacts with both nicked DNA and the DNA sliding clamps, PCNA and hRad9-hRad1-hHus1.
abstract: The participation of the DNA ligase (hLigI) encoded by the human LIG1 gene in DNA replication and repair is mediated by an interaction with proliferating cell nuclear antigen (PCNA), a homotrimeric DNA sliding clamp. Interestingly, the catalytic fragment of hLigI encircles a DNA nick forming a ring that is similar in size and shape to the PCNA ring. Here we show that the DNA binding domain (DBD) within the hLigI catalytic fragment interacts with both PCNA and the heterotrimeric cell-cycle checkpoint clamp, hRad9-hRad1-hHus1 (9-1-1). The DBD preferentially binds to trimeric PCNA and the hRad1 subunit of 9-1-1. Unlike the majority of PCNA interacting proteins, the DBD does not interact with the interdomain connector loop region of PCNA but instead appears to interact with regions adjacent to the intersubunit interfaces within the PCNA trimer. Notably, the DBD not only binds specifically to DNA nicks but also mediates the formation of DNA protein complexes with PCNA. Based on these results, we suggest that the interface between the DBD and PCNA acts as a pivot facilitating the transition of the hLigI catalytic region fragment from an extended conformation to a ring structure when it engages a DNA nick.
authors.affiliation: Radiation Oncology Research Laboratory and Marlene & Stewart Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Song
authors.forename: John M
authors.initials: JM
authors.lastname: Pascal
authors.forename: Tom
authors.initials: T
authors.lastname: Ellenberger
authors.forename: Alan E
authors.initials: AE
authors.lastname: Tomkinson
mesh_terms: D020134:Catalytic Domain; D018797:Cell Cycle Proteins / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D004249:DNA Damage; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000737:chemistry* / Q000378:metabolism*; D005092:Exonucleases / Q000378:metabolism*; D006801:Humans; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D055503:Protein Multimerization; D021122:Protein Subunits / Q000378:metabolism
chemical_list: D018797:Cell Cycle Proteins; C462949:HUS1B protein, human; C000607892:LIG1 protein, human; D018809:Proliferating Cell Nuclear Antigen; D021122:Protein Subunits; C069930:rad9 protein; D004247:DNA; D005092:Exonucleases; C113574:Rad1 protein, human; D011088:DNA Ligases; D000072481:DNA Ligase ATP