Structural and torsional properties of the RAD51-dsDNA nucleoprotein filament.

Publisher: Oxford University Press Country of Publication: England NLM ID: 0411011 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1362-4962 (Electronic) Linking ISSN: 03051048 NLM ISO Abbreviation: Nucleic Acids Res Subsets: MEDLINE

Publication: 1992- : Oxford : Oxford University Press
Original Publication: London, Information Retrieval ltd.

DNA/*chemistry ; Rad51 Recombinase/*chemistry; DNA/metabolism ; Humans ; Nucleoproteins/chemistry ; Nucleoproteins/metabolism ; Rad51 Recombinase/metabolism ; Torsion, Mechanical

Human RAD51 is a key protein in the repair of DNA by homologous recombination. Its assembly onto DNA, which induces changes in DNA structure, results in the formation of a nucleoprotein filament that forms the basis of strand exchange. Here, we determine the structural and mechanical properties of RAD51-dsDNA filaments. Our measurements use two recently developed magnetic tweezers assays, freely orbiting magnetic tweezers and magnetic torque tweezers, designed to measure the twist and torque of individual molecules. By directly monitoring changes in DNA twist on RAD51 binding, we determine the unwinding angle per RAD51 monomer to be 45°, in quantitative agreement with that of its bacterial homolog, RecA. Measurements of the torque that is built up when RAD51-dsDNA filaments are twisted show that under conditions that suppress ATP hydrolysis the torsional persistence length of the RAD51-dsDNA filament exceeds that of its RecA counterpart by a factor of three. Examination of the filament's torsional stiffness for different combinations of divalent ions and nucleotide cofactors reveals that the Ca(2+) ion, apart from suppressing ATPase activity, plays a key role in increasing the torsional stiffness of the filament. These quantitative measurements of RAD51-imposed DNA distortions and accumulated mechanical stress suggest a finely tuned interplay between chemical and mechanical interactions within the RAD51 nucleoprotein filament.

Annu Rev Biochem. 2008;77:229-57. (PMID: 18275380)
Phys Chem Chem Phys. 2009 Jun 28;11(24):4800-3. (PMID: 19506753)
Curr Protein Pept Sci. 2004 Apr;5(2):73-9. (PMID: 15078218)
DNA Repair (Amst). 2010 Dec 10;9(12):1264-72. (PMID: 20971042)
J Biol Chem. 2005 Jul 22;280(29):26886-95. (PMID: 15917244)
Nucleic Acids Res. 2005 Jun 08;33(10):3292-302. (PMID: 15944450)
Proc Natl Acad Sci U S A. 2004 Jul 6;101(27):9988-93. (PMID: 15226506)
Structure. 2007 May;15(5):599-609. (PMID: 17502105)
Proc Natl Acad Sci U S A. 1999 Aug 31;96(18):10109-14. (PMID: 10468570)
Mol Cell. 2004 Aug 13;15(3):423-35. (PMID: 15304222)
Nucleic Acids Res. 2012 Dec;40(22):11769-76. (PMID: 23180779)
Nano Lett. 2011 Dec 14;11(12):5558-63. (PMID: 22047401)
Science. 1993 Mar 26;259(5103):1896-9. (PMID: 8456314)
Nucleic Acids Res. 2007;35(21):7171-87. (PMID: 17947332)
Proc Natl Acad Sci U S A. 1999 Jul 6;96(14):7916-21. (PMID: 10393922)
Nature. 1992 Jan 23;355(6358):318-25. (PMID: 1731246)
Mol Cell. 2012 Jun 8;46(5):616-24. (PMID: 22560720)
J Biol Chem. 2006 Dec 22;281(51):39380-7. (PMID: 17050545)
Nat Methods. 2010 Dec;7(12):977-80. (PMID: 20953173)
Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8419-24. (PMID: 11459984)
Nucleic Acids Res. 2012 Jun;40(11):4904-13. (PMID: 22362735)
Proc Natl Acad Sci U S A. 2009 Nov 17;106(46):19239-44. (PMID: 19884492)
Proc Natl Acad Sci U S A. 2009 Aug 11;106(32):13248-53. (PMID: 19587234)
Trends Biochem Sci. 1998 Jul;23(7):247-51. (PMID: 9697414)
Nature. 2008 May 22;453(7194):489-4. (PMID: 18497818)
EMBO J. 1994 Dec 1;13(23):5764-71. (PMID: 7988572)
Biophys J. 1999 Jan;76(1 Pt 1):409-13. (PMID: 9876152)
Methods Mol Biol. 2009;582:71-89. (PMID: 19763943)
Biophys J. 2010 Aug 9;99(4):1292-302. (PMID: 20713015)
Nature. 2012 Feb 08;482(7385):423-7. (PMID: 22318518)
Nucleic Acids Res. 2007;35(17):5646-57. (PMID: 17709342)
Rev Sci Instrum. 2011 Oct;82(10):103707. (PMID: 22047303)
J Mol Biol. 2001 Jun 8;309(3):539-42. (PMID: 11397077)
Phys Rev Lett. 2008 Apr 11;100(14):148301. (PMID: 18518075)
Elife. 2012 Dec 13;1:e00067. (PMID: 23240082)
Nature. 2009 Feb 5;457(7230):745-8. (PMID: 19060884)
Curr Opin Struct Biol. 2012 Jun;22(3):304-12. (PMID: 22658779)
Nat Commun. 2011 Aug 23;2:439. (PMID: 21863006)
Nano Lett. 2012 Jul 11;12(7):3634-9. (PMID: 22642488)
Phys Rev Lett. 2009 Feb 20;102(7):078301. (PMID: 19257716)
Nat Struct Mol Biol. 2004 Aug;11(8):791-6. (PMID: 15235592)
J Biol Chem. 2005 Jan 7;280(1):722-8. (PMID: 15537659)
Nano Lett. 2009 Apr;9(4):1720-5. (PMID: 19301859)

0 (Nucleoproteins)
9007-49-2 (DNA)
EC 2.7.7.- (RAD51 protein, human)
EC 2.7.7.- (Rad51 Recombinase)

Date Created: 20130525 Date Completed: 20131104 Latest Revision: 20211021

20240829

PMC3737536

10.1093/nar/gkt425

23703213