Tags

Type your tag names separated by a space and hit enter

Protein dynamics during presynaptic-complex assembly on individual single-stranded DNA molecules.
Nat Struct Mol Biol. 2014 Oct; 21(10):893-900.NS

Abstract

Homologous recombination is a conserved pathway for repairing double-stranded breaks, which are processed to yield single-stranded DNA overhangs that serve as platforms for presynaptic-complex assembly. Here we use single-molecule imaging to reveal the interplay between Saccharomyces cerevisiae RPA, Rad52 and Rad51 during presynaptic-complex assembly. We show that Rad52 binds RPA-ssDNA and suppresses RPA turnover, highlighting an unanticipated regulatory influence on protein dynamics. Rad51 binding extends the ssDNA, and Rad52-RPA clusters remain interspersed along the presynaptic complex. These clusters promote additional binding of RPA and Rad52. Our work illustrates the spatial and temporal progression of the association of RPA and Rad52 with the presynaptic complex and reveals a new RPA-Rad52-Rad51-ssDNA intermediate, with implications for how the activities of Rad52 and RPA are coordinated with Rad51 during the later stages of recombination.

Authors+Show Affiliations

Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York, USA.Department of Biological Sciences, Columbia University, New York, New York, USA.Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut, USA.Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut, USA.Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut, USA.1] Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York, USA. [2] Howard Hughes Medical Institute, Columbia University, New York, New York, USA.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

25195049

Citation

Gibb, Bryan, et al. "Protein Dynamics During Presynaptic-complex Assembly On Individual Single-stranded DNA Molecules." Nature Structural & Molecular Biology, vol. 21, no. 10, 2014, pp. 893-900.
Gibb B, Ye LF, Kwon Y, et al. Protein dynamics during presynaptic-complex assembly on individual single-stranded DNA molecules. Nat Struct Mol Biol. 2014;21(10):893-900.
Gibb, B., Ye, L. F., Kwon, Y., Niu, H., Sung, P., & Greene, E. C. (2014). Protein dynamics during presynaptic-complex assembly on individual single-stranded DNA molecules. Nature Structural & Molecular Biology, 21(10), 893-900. https://doi.org/10.1038/nsmb.2886
Gibb B, et al. Protein Dynamics During Presynaptic-complex Assembly On Individual Single-stranded DNA Molecules. Nat Struct Mol Biol. 2014;21(10):893-900. PubMed PMID: 25195049.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Protein dynamics during presynaptic-complex assembly on individual single-stranded DNA molecules. AU - Gibb,Bryan, AU - Ye,Ling F, AU - Kwon,YoungHo, AU - Niu,Hengyao, AU - Sung,Patrick, AU - Greene,Eric C, Y1 - 2014/09/07/ PY - 2014/03/05/received PY - 2014/08/12/accepted PY - 2014/9/8/entrez PY - 2014/9/10/pubmed PY - 2015/1/8/medline SP - 893 EP - 900 JF - Nature structural & molecular biology JO - Nat Struct Mol Biol VL - 21 IS - 10 N2 - Homologous recombination is a conserved pathway for repairing double-stranded breaks, which are processed to yield single-stranded DNA overhangs that serve as platforms for presynaptic-complex assembly. Here we use single-molecule imaging to reveal the interplay between Saccharomyces cerevisiae RPA, Rad52 and Rad51 during presynaptic-complex assembly. We show that Rad52 binds RPA-ssDNA and suppresses RPA turnover, highlighting an unanticipated regulatory influence on protein dynamics. Rad51 binding extends the ssDNA, and Rad52-RPA clusters remain interspersed along the presynaptic complex. These clusters promote additional binding of RPA and Rad52. Our work illustrates the spatial and temporal progression of the association of RPA and Rad52 with the presynaptic complex and reveals a new RPA-Rad52-Rad51-ssDNA intermediate, with implications for how the activities of Rad52 and RPA are coordinated with Rad51 during the later stages of recombination. SN - 1545-9985 UR - https://www.unboundmedicine.com/medline/citation/25195049/Protein_dynamics_during_presynaptic_complex_assembly_on_individual_single_stranded_DNA_molecules_ L2 - https://doi.org/10.1038/nsmb.2886 DB - PRIME DP - Unbound Medicine ER -