7EP8

Crystal structure of PCNA from Neurospora crassa

7EP8 の概要

• エントリーDOI: 10.2210/pdb7ep8/pdb
• 分子名称: Proliferating cell nuclear antigen (2 entities in total)
• 機能のキーワード: dna sliding clamp, dna replication, dna repair, dna binding protein
• 由来する生物種: Neurospora crassa OR74A
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 28301.19

構造登録者

Sundaram, R.,Vasudevan, D. (登録日: 2021-04-26, 公開日: 2021-06-30, 最終更新日: 2023-11-29)

主引用文献

Kumari, P.,Sundaram, R.,Manohar, K.,Vasudevan, D.,Acharya, N.
Interdomain connecting loop and J loop structures determine cross-species compatibility of PCNA.
J.Biol.Chem., 297:100911-100911, 2021

PubMed Abstract: Eukaryotic proliferating cell nuclear antigen (PCNA) plays an essential role in orchestrating the assembly of the replisome complex, stimulating processive DNA synthesis, and recruiting other regulatory proteins during the DNA damage response. PCNA and its binding partner network are relatively conserved in eukaryotes, and it exhibits extraordinary structural similarity across species. However, despite this structural similarity, the PCNA of a given species is rarely functional in heterologous systems. In this report, we determined the X-ray crystal structure of Neurospora crassa PCNA (NcPCNA) and compared its structure-function relationship with other available PCNA studies to understand this cross-species incompatibility. We found two regions, the interdomain connecting loop (IDCL) and J loop structures, vary significantly among PCNAs. In particular, the J loop deviates in NcPCNA from that in Saccharomyces cerevisiae PCNA (ScPCNA) by 7 Å. Differences in the IDCL structures result in varied binding affinities of PCNAs for the subunit Pol32 of DNA polymerase delta and for T2-amino alcohol, a small-molecule inhibitor of human PCNA. To validate that these structural differences are accountable for functional incompatibility in S. cerevisiae, we generated NcPCNA mutants mimicking IDCL and J loop structures of ScPCNA. Our genetic analyses suggested that NcPCNA mutants are fully functional in S. cerevisiae. The susceptibility of the strains harboring ScPCNA mimics of NcPCNA to various genotoxic agents was similar to that in yeast cells expressing ScPCNA. Taken together, we conclude that in addition to the overall architecture of PCNA, structures of the IDCL and J loop of PCNA are critical determinants of interspecies functional compatibility.

PubMed: 34175309
DOI: 10.1016/j.jbc.2021.100911

実験手法

X-RAY DIFFRACTION (1.95 Å)