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An interaction network in the polymerase active site is a prerequisite for Watson-Crick base pairing in Pol γ

Journal Article · · Science Advances
 [1];  [2];  [3];  [4];  [3];  [1]
  1. University of Texas Medical Branch, Galveston, TX (United States)
  2. University of Texas Medical Branch, Galveston, TX (United States); Baylor College of Medicine, Houston, TX (United States)
  3. Univ. of Texas at Dallas, Richardson, TX (United States)
  4. University of Texas Medical Branch, Galveston, TX (United States); University of Texas Health Science Center at Houston, TX (United States)
+ Show Author Affiliations
The replication accuracy of DNA polymerase gamma (Pol γ) is essential for mitochondrial genome integrity. Mutation of human Pol γ arginine-853 has been linked to neurological diseases. Although not a catalytic residue, Pol γ arginine-853 mutants are void of polymerase activity. To identify the structural basis for the disease, we determined a crystal structure of the Pol γ mutant ternary complex with correct incoming nucleotide 2'-deoxycytidine 5'-triphosphate (dCTP). Opposite to the wild type that undergoes open-to-closed conformational changes when bound to a correct nucleotide that is essential for forming a catalytically competent active site, the mutant complex failed to undergo the conformational change, and the dCTP did not base pair with its Watson-Crick complementary templating residue. Our studies revealed that arginine-853 coordinates an interaction network that aligns the 3'-end of primer and dCTP with the catalytic residues. Disruption of the network precludes the formation of Watson-Crick base pairing and closing of the active site, resulting in an inactive polymerase.
Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
National Institutes of Health (NIH); National Science Foundation (NSF); USDOE Office of Science (SC), Basic Energy Sciences (BES)
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
2469664
Journal Information:
Science Advances, Journal Name: Science Advances Journal Issue: 21 Vol. 10; ISSN 2375-2548
Publisher:
AAASCopyright Statement
Country of Publication:
United States
Language:
English

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