Journal article
Analysis of the Activities of RAD54, a SWI2/SNF2 Protein, Using a Specific Small-molecule Inhibitor
The Journal of biological chemistry, v 288(44), pp 31567-31580
01 Nov 2013
PMID: 24043618
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Background:
RAD54 performs branch migration and stimulation of RAD51 DNA strand exchange.
Results:
Streptonigrin inhibits branch migration through specific binding to RAD54 but spares stimulation of RAD51-mediated DNA strand exchange.
Conclusion:
RAD54 ATPase activity is relatively less important in the stimulation of DNA pairing than in branch migration.
Significance:
Small-molecule inhibitors are instrumental to study the mechanisms of RAD54 activities.
RAD54, an important homologous recombination protein, is a member of the SWI2/SNF2 family of ATPase-dependent DNA translocases.
In vitro
, RAD54 stimulates RAD51-mediated DNA strand exchange and promotes branch migration of Holliday junctions. It is thought that an ATPase-dependent DNA translocation is required for both of these RAD54 activities. Here we identified, by high-throughput screening, a specific RAD54 inhibitor, streptonigrin (SN), and used it to investigate the mechanisms of RAD54 activities. We found that SN specifically targets the RAD54 ATPase, but not DNA binding, through direct interaction with RAD54 and generation of reactive oxygen species. Consistent with the dependence of branch migration (BM) on the ATPase-dependent DNA translocation of RAD54, SN inhibited RAD54 BM. Surprisingly, the ability of RAD54 to stimulate RAD51 DNA strand exchange was not significantly affected by SN, indicating a relatively smaller role of RAD54 DNA translocation in this process. Thus, the use of SN enabled us to identify important differences in the effect of the RAD54 ATPase and DNA translocation on two major activities of RAD54, BM of Holliday junctions and stimulation of DNA pairing.
Metrics
Details
- Title
- Analysis of the Activities of RAD54, a SWI2/SNF2 Protein, Using a Specific Small-molecule Inhibitor
- Creators
- Julianna S Deakyne - From theFei Huang - From theJoseph Negri - theNicola Tolliday - theSimon Cocklin - From theAlexander V Mazin - From the
- Publication Details
- The Journal of biological chemistry, v 288(44), pp 31567-31580
- Publisher
- American Society for Biochemistry and Molecular Biology; 9650 Rockville Pike, Bethesda, MD 20814, U.S.A
- Grant note
- CA100839; DA033981; MH097512; 1R21AI087388 / National Institutes of Health
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:000330596200016
- Scopus ID
- 2-s2.0-84887064326
- Other Identifier
- 991014878407604721
UN Sustainable Development Goals (SDGs)
This publication has contributed to the advancement of the following goals:
InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Biochemistry & Molecular Biology