7lfq
From Proteopedia
Pyrococcus RNA ligase
Structural highlights
FunctionRTCB_PYRHO tRNA-splicing ligase that acts by directly joining spliced tRNA halves to mature-sized tRNAs by incorporating the precursor-derived splice junction phosphate into the mature tRNA as a canonical 3',5'-phosphodiester. May act as a RNA ligase with broad substrate specificity, and may function toward other RNAs (By similarity). Publication Abstract from PubMedRtcB enzymes comprise a widely distributed family of manganese- and GTP-dependent RNA repair enzymes that join 2',3'-cyclic phosphate ends to 5'-OH ends via RtcB-(histidinyl-N)-GMP, RNA 3'-phosphate, and RNA3'pp5'G intermediates. RtcB can ligate either 5'-OH RNA or 5'-OH DNA strands in vitro. The nucleic acid contacts of RtcB are uncharted. Here we report a 2.7 A crystal structure of Pyrococcus horikoshii RtcB in complex with a 6-mer 5'-OH DNA oligonucleotide (HO)A(1)pT(2)pG(3)pT(4)pC(5)pC(6), which reveals enzymic contacts of Asn202 to the terminal 5'-OH nucleophile; Arg238 to the A(1)pT(2) and T(2)pG(3) phosphates; Arg190 and Gln194 to the T(2)pG(3) phosphate; and an Arg190 pi-cation interaction with the G3 nucleobase. The structural insights affirm functional studies of E. coli RtcB that implicated the conserved counterpart of Arg238 in engagement of the 5'-OH strand for ligation. The essential active site Cys98 that coordinates two manganese ions is oxidized to cysteine sulfonic acid in our structure, raising the prospect that RtcB activity might be sensitive to modulation during oxidative stress. Structure of 3'-PO(4)/5'-OH RNA ligase RtcB in complex with a 5'-OH oligonucleotide.,Banerjee A, Goldgur Y, Shuman S RNA. 2021 Feb 22;27(5):584-90. doi: 10.1261/rna.078692.121. PMID:33619169[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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