5X4I

Pyrococcus furiosus RecJ (D83A, Mn-soaking)

Summary for 5X4I

• Entry DOI: 10.2210/pdb5x4i/pdb
• Related: 5X4H 5X4J 5X4K
• Descriptor: Uncharacterized protein, MANGANESE (II) ION, CHLORIDE ION, ... (4 entities in total)
• Functional Keywords: archaeal recj, cmg interaction domain, nuclease activity, gins, interaction, hydrolase
• Biological source: Pyrococcus furiosus
• Total number of polymer chains: 1
• Total formula weight: 56470.32

Authors

Li, M.J.,Yi, G.S.,Yu, F.,Zhou, H.,Chen, J.N.,Xu, C.Y.,Wang, F.P.,Xiao, X.,He, J.H.,Liu, X.P. (deposition date: 2017-02-13, release date: 2018-02-14, Last modification date: 2024-10-09)

Primary citation

Li, M.J.,Yi, G.S.,Yu, F.,Zhou, H.,Chen, J.N.,Xu, C.Y.,Wang, F.P.,Xiao, X.,He, J.H.,Liu, X.P.
The crystal structure of Pyrococcus furiosus RecJ implicates it as an ancestor of eukaryotic Cdc45.
Nucleic Acids Res., 45:12551-12564, 2017

PubMed Abstract: RecJ nucleases specifically degrade single-stranded (ss) DNA in the 5' to 3' direction. Archaeal RecJ is different from bacterial RecJ in sequence, domain organization, and substrate specificity. The RecJ from archaea Pyrococcus furiosus (PfuRecJ) also hydrolyzes RNA strands in the 3' to 5' direction. Like eukaryotic Cdc45 protein, archaeal RecJ forms a complex with MCM helicase and GINS. Here, we report the crystal structures of PfuRecJ and the complex of PfuRecJ and two CMPs. PfuRecJ bind one or two divalent metal ions in its crystal structure. A channel consisting of several positively charged residues is identified in the complex structure, and might be responsible for binding substrate ssDNA and/or releasing single nucleotide products. The deletion of the complex interaction domain (CID) increases the values of kcat/Km of 5' exonuclease activity on ssDNA and 3' exonuclease activity on ssRNA by 5- and 4-fold, respectively, indicating that the CID functions as a regulator of enzymatic activity. The DHH domain of PfuRecJ interacts with the C-terminal beta-sheet domain of the GINS51 subunit in the tetrameric GINS complex. The relationship of archaeal and bacterial RecJs, as well as eukaryotic Cdc45, is discussed based on biochemical and structural results.

PubMed: 30053256
DOI: 10.1093/nar/gkx887

Experimental method

X-RAY DIFFRACTION (2.092 Å)