title: P50, the small subunit of DNA polymerase delta, is required for mediation of the interaction of polymerase delta subassemblies with PCNA.
abstract: Mammalian DNA polymerase δ (pol δ), a four-subunit enzyme, plays a crucial and versatile role in DNA replication and various DNA repair processes. Its function as a chromosomal DNA polymerase is dependent on the association with proliferating cell nuclear antigen (PCNA) which functions as a molecular sliding clamp. All four of the pol δ subunits (p125, p50, p68, and p12) have been reported to bind to PCNA. However, the identity of the subunit of pol δ that directly interacts with PCNA and is therefore primarily responsible for the processivity of the enzyme still remains controversial. Previous model for the network of protein-protein interactions of the pol δ-PCNA complex showed that pol δ might be able to interact with a single molecule of PCNA homotrimer through its three subunits, p125, p68, and p12 in which the p50 was not included in. Here, we have confirmed that the small subunit p50 of human pol δ truthfully interacts with PCNA by the use of far-Western analysis, quantitative ELISA assay, and subcellular co-localization. P50 is required for mediation of the interaction between pol δ subassemblies and PCNA homotrimer. Thus, pol δ interacts with PCNA via its four subunits.
authors.affiliation: Institute of Life Sciences, Jiangsu University, Zhenjiang, Jiangsu, People's Republic of China.
authors.forename: Yujue
authors.initials: Y
authors.lastname: Wang
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.forename: Huiqing
authors.initials: H
authors.lastname: Chen
authors.forename: Xiao
authors.initials: X
authors.lastname: Li
authors.forename: Weijun
authors.initials: W
authors.lastname: Mai
authors.forename: Keping
authors.initials: K
authors.lastname: Chen
authors.forename: Sufang
authors.initials: S
authors.lastname: Zhang
authors.forename: Ernest Y C
authors.initials: EY
authors.lastname: Lee
authors.forename: Marietta Y W T
authors.initials: MY
authors.lastname: Lee
authors.forename: Yajing
authors.initials: Y
authors.lastname: Zhou
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D004258:DNA Polymerase III / Q000737:chemistry / Q000276:immunology / Q000502:physiology*; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D018809:Proliferating Cell Nuclear Antigen / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D017931:DNA Primers; D018809:Proliferating Cell Nuclear Antigen; D004258:DNA Polymerase III
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