Title |
Direct imaging of human Rad51 nucleoprotein dynamics on individual DNA molecules
|
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Published in |
Proceedings of the National Academy of Sciences of the United States of America, January 2009
|
DOI | 10.1073/pnas.0811965106 |
Pubmed ID | |
Authors |
Jovencio Hilario, Ichiro Amitani, Ronald J. Baskin, Stephen C. Kowalczykowski |
Abstract |
Rad51 protein (Rad51) is central to recombinational repair of double-strand DNA breaks. It polymerizes onto DNA and promotes strand exchange between homologous chromosomes. We visualized the real-time assembly and disassembly of human Rad51 nucleoprotein filaments on double-stranded DNA by single-molecule fluorescence microscopy. Rad51 assembly extends the DNA by approximately 65%. Nucleoprotein filament formation occurs via rapid nucleation followed by growth from these nuclei. Growth does not continue indefinitely, however, and nucleoprotein filaments terminate when approximately 2 mum in length. The dependence of nascent filament formation on Rad51 concentration suggests that 2-3 Rad51 monomers are involved in nucleation. Rad51 nucleoprotein filaments are stable and remain extended when ATP hydrolysis is prevented; however, when permitted, filaments decrease in length as a result of conversion to ADP-bound nucleoprotein complexes and partial protein dissociation. Dissociation of Rad51 from dsDNA is slow and incomplete, thereby rationalizing the need for other proteins that facilitate disassembly. |
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Geographical breakdown
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United Kingdom | 2 | 1% |
Japan | 2 | 1% |
Italy | 1 | <1% |
Vietnam | 1 | <1% |
Korea, Republic of | 1 | <1% |
Iran, Islamic Republic of | 1 | <1% |
Netherlands | 1 | <1% |
France | 1 | <1% |
Other | 1 | <1% |
Unknown | 168 | 92% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 48 | 26% |
Researcher | 43 | 24% |
Student > Master | 18 | 10% |
Student > Bachelor | 14 | 8% |
Professor | 10 | 5% |
Other | 34 | 19% |
Unknown | 15 | 8% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 66 | 36% |
Biochemistry, Genetics and Molecular Biology | 59 | 32% |
Physics and Astronomy | 19 | 10% |
Chemistry | 12 | 7% |
Unspecified | 3 | 2% |
Other | 9 | 5% |
Unknown | 14 | 8% |