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The ATPase mechanism of UvrA2 reveals the distinct roles of proximal and distal ATPase sites in nucleotide excision repair

Journal Article · · Nucleic Acids Research
DOI:https://doi.org/10.1093/nar/gkz180· OSTI ID:1625577
 [1];  [2];  [3];  [4];  [5]
  1. Wesleyan Univ., Middletown, CT (United States). Dept. of Molecular Biology and Biochemistry; DOE/OSTI
  2. City College of New York, NY (United States). Dept. of Chemistry and Biochemistry; City College of New York, NY (United States). The Graduate Center. Ph.D. Program in Biochemistry
  3. City College of New York, NY (United States). Dept. of Chemistry and Biochemistry; Hunter College High School, New York, NY (United States)
  4. City College of New York, NY (United States). Dept. of Chemistry and Biochemistry; City College of New York, NY (United States). The Graduate Center. Ph.D. Program in Biochemistry; City College of New York, NY (United States). The Graduate Center. Ph.D. Programs in Chemistry and Biology
  5. Wesleyan Univ., Middletown, CT (United States). Dept. of Molecular Biology and Biochemistry
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The UvrA2 dimer finds lesions in DNA and initiates nucleotide excision repair. Each UvrA monomer contains two essential ATPase sites: proximal (P) and distal (D). The manner whereby their activities enable UvrA2 damage sensing and response remains to be clarified. We report three key findings from the first pre-steady state kinetic analysis of each site. Absent DNA, a P2ATP-D2ADP species accumulates when the low-affinity proximal sites bind ATP and enable rapid ATP hydrolysis and phosphate release by the high affinity distal sites, and ADP release limits catalytic turnover. Native DNA stimulates ATP hydrolysis by all four sites, causing UvrA2 to transition through a different species, P2ADP-D2ADP. Lesion-containing DNA changes the mechanism again, suppressing ATP hydrolysis by the proximal sites while distal sites cycle through hydrolysis and ADP release, to populate proximal ATP-bound species, P2ATP-Dempty and P2ATPD2ATP. Thus, damaged and native DNA trigger distinct ATPase site activities, which could explain why UvrA2 forms stable complexes with UvrB on damaged DNA compared with weaker, more dynamic complexes on native DNA. Such specific coupling between the DNA substrate and the ATPase mechanism of each site provides new insights into how UvrA2 utilizes ATP for lesion search, recognition and repair.
Research Organization:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division
Grant/Contract Number:
SC0012704
OSTI ID:
1625577
Alternate ID(s):
OSTI ID: 1900833
Journal Information:
Nucleic Acids Research, Journal Name: Nucleic Acids Research Journal Issue: 8 Vol. 47; ISSN 0305-1048
Publisher:
Oxford University PressCopyright Statement
Country of Publication:
United States
Language:
English

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