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

Autor: Memie Osuga, Silas Hartley, Brandon C Case, David Jeruzalmi, Manju M. Hingorani
Rok vydání: 2019
Předmět:
DNA
Bacterial
Models
Molecular
DNA Repair
DNA damage
DNA repair
ATPase
Genetic Vectors
Gene Expression
Biology
Protein Structure
Secondary
Substrate Specificity
Geobacillus stearothermophilus
03 medical and health sciences
chemistry.chemical_compound
Adenosine Triphosphate
0302 clinical medicine
Bacterial Proteins
ATP hydrolysis
Escherichia coli
Genetics
Protein Interaction Domains and Motifs
Thermotoga maritima
ortho-Aminobenzoates
Nucleotide
Amino Acid Sequence
Cloning
Molecular
Binding site
030304 developmental biology
chemistry.chemical_classification
0303 health sciences
Binding Sites
Endodeoxyribonucleases
Nucleic Acid Enzymes
Escherichia coli Proteins
Recombinant Proteins
Kinetics
chemistry
Structural Homology
Protein
Biophysics
biology.protein
Thermodynamics
Protein Multimerization
Sequence Alignment
030217 neurology & neurosurgery
DNA
DNA Damage
Protein Binding
Nucleotide excision repair
Zdroj: Nucleic Acids Research
ISSN: 1362-4962
0305-1048
DOI: 10.1093/nar/gkz180
Popis: 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 P2ATP-D2ATP. 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.
Databáze: OpenAIRE
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