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Title: Atomic structures of the RNA end-healing 5′-OH kinase and 2′,3′-cyclic phosphodiesterase domains of fungal tRNA ligase: conformational switches in the kinase upon binding of the GTP phosphate donor

Journal Article · · Nucleic Acids Research
DOI:https://doi.org/10.1093/nar/gkz1049· OSTI ID:1607550
 [1];  [2];  [3];  [1]
  1. Molecular Biology Program, Sloan-Kettering Institute, New York, NY 10065, USA
  2. Structural Biology Program, Sloan-Kettering Institute, New York, NY 10065, USA
  3. Microbiology and Immunology Department, Weill Cornell Medical College, New York, NY 10065, USA
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Fungal tRNA ligase (Trl1) rectifies RNA breaks with 2',3'-cyclic-PO4 and 5'-OH termini. Trl1 consists of three catalytic modules: an N-terminal ligase (LIG) domain; a central polynucleotide kinase (KIN) domain; and a C-terminal cyclic phosphodiesterase (CPD) domain. Trl1 enzymes found in all human fungal pathogens are untapped targets for antifungal drug discovery. Here we report a 1.9 Å crystal structure of Trl1 KIN-CPD from the pathogenic fungus Candida albicans, which adopts an extended conformation in which separate KIN and CPD domains are connected by an unstructured linker. CPD belongs to the 2H phosphotransferase superfamily by dint of its conserved central concave β sheet and interactions of its dual HxT motif histidines and threonines with phosphate in the active site. Additional active site motifs conserved among the fungal CPD clade of 2H enzymes are identified. We present structures of the Candida Trl1 KIN domain at 1.5 to 2.0 Å resolution—as apoenzyme and in complexes with GTP•Mg2+, IDP•PO4, and dGDP•PO4—that highlight conformational switches in the G-loop (which recognizes the guanine base) and lid-loop (poised over the nucleotide phosphates) that accompany nucleotide binding.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
USDOE; National Institutes of Health (NIH); National Cancer Institute (NCI); German Research Foundation (DFG)
Grant/Contract Number:
AC02-06CH11357; R35-GM126945; 394320208; P30-CA008748; P41GM103403; HEI-S10RR029205
OSTI ID:
1607550
Alternate ID(s):
OSTI ID: 1578227
Journal Information:
Nucleic Acids Research, Journal Name: Nucleic Acids Research; ISSN 0305-1048
Publisher:
Oxford University PressCopyright Statement
Country of Publication:
United Kingdom
Language:
English
Citation Metrics:
Cited by: 4 works
Citation information provided by
Web of Science

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