The N-terminal domain of the human Rad51 protein binds DNA: structure and a DNA binding surface as revealed by NMR - PubMed (original) (raw)

Comparative Study

. 1999 Jul 9;290(2):495-504.

doi: 10.1006/jmbi.1999.2904.

Affiliations

• PMID: 10390347
• DOI: 10.1006/jmbi.1999.2904

Comparative Study

The N-terminal domain of the human Rad51 protein binds DNA: structure and a DNA binding surface as revealed by NMR

H Aihara et al. J Mol Biol. 1999.

Abstract

Human Rad51 protein (HsRad51) is a homolog of Escherichia coli RecA protein, and functions in DNA repair and recombination. In higher eukaryotes, Rad51 protein is essential for cell viability. The N-terminal region of HsRad51 is highly conserved among eukaryotic Rad51 proteins but is absent from RecA, suggesting a Rad51-specific function for this region. Here, we have determined the structure of the N-terminal part of HsRad51 by NMR spectroscopy. The N-terminal region forms a compact domain consisting of five short helices, which shares structural similarity with a domain of endonuclease III, a DNA repair enzyme of E. coli. NMR experiments did not support the involvement of the N-terminal domain in HsRad51-HsBrca2 interaction or the self-association of HsRad51 as proposed by previous studies. However, NMR tiration experiments demonstrated a physical interaction of the domain with DNA, and allowed mapping of the DNA binding surface. Mutation analysis showed that the DNA binding surface is essential for double-stranded and single-stranded DNA binding of HsRad51. Our results suggest the presence of a DNA binding site on the outside surface of the HsRad51 filament and provide a possible explanation for the regulation of DNA binding by phosphorylation within the N-terminal domain.

Copyright 1999 Academic Press.

PubMed Disclaimer

Similar articles

• Human Rad51 amino acid residues required for Rad52 binding.
  Kurumizaka H, Aihara H, Kagawa W, Shibata T, Yokoyama S. Kurumizaka H, et al. J Mol Biol. 1999 Aug 20;291(3):537-48. doi: 10.1006/jmbi.1999.2950. J Mol Biol. 1999. PMID: 10448035
• An interaction between a specified surface of the C-terminal domain of RecA protein and double-stranded DNA for homologous pairing.
  Aihara H, Ito Y, Kurumizaka H, Terada T, Yokoyama S, Shibata T. Aihara H, et al. J Mol Biol. 1997 Nov 28;274(2):213-21. doi: 10.1006/jmbi.1997.1403. J Mol Biol. 1997. PMID: 9398528
• Gly-103 in the N-terminal domain of Saccharomyces cerevisiae Rad51 protein is critical for DNA binding.
  Zhang XP, Lee KI, Solinger JA, Kiianitsa K, Heyer WD. Zhang XP, et al. J Biol Chem. 2005 Jul 15;280(28):26303-11. doi: 10.1074/jbc.M503244200. Epub 2005 May 21. J Biol Chem. 2005. PMID: 15908697
• Homologous genetic recombination as an intrinsic dynamic property of a DNA structure induced by RecA/Rad51-family proteins: a possible advantage of DNA over RNA as genomic material.
  Shibata T, Nishinaka T, Mikawa T, Aihara H, Kurumizaka H, Yokoyama S, Ito Y. Shibata T, et al. Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8425-32. doi: 10.1073/pnas.111005198. Proc Natl Acad Sci U S A. 2001. PMID: 11459985 Free PMC article. Review.
• Mammalian Rad51 protein: a RecA homologue with pleiotropic functions.
  Vispé S, Defais M. Vispé S, et al. Biochimie. 1997 Oct;79(9-10):587-92. doi: 10.1016/s0300-9084(97)82007-x. Biochimie. 1997. PMID: 9466696 Review.

Cited by

• Molecular cloning and characterization of RAD51-like genes from Arabidopsis thaliana.
  Osakabe K, Yoshioka T, Ichikawa H, Toki S. Osakabe K, et al. Plant Mol Biol. 2002 Sep;50(1):71-81. doi: 10.1023/a:1016047231597. Plant Mol Biol. 2002. PMID: 12139010
• Rad51 protein from the thermotolerant yeast Pichia angusta as a typical but thermodependent member of the Rad51 family.
  Shalguev VI, Kil YV, Yurchenko LV, Namsaraev EA, Lanzov VA. Shalguev VI, et al. Eukaryot Cell. 2004 Dec;3(6):1567-73. doi: 10.1128/EC.3.6.1567-1573.2004. Eukaryot Cell. 2004. PMID: 15590830 Free PMC article.
• Structural and functional analyses of five conserved positively charged residues in the L1 and N-terminal DNA binding motifs of archaeal RADA protein.
  Chen LT, Ko TP, Chang YW, Lin KA, Wang AH, Wang TF. Chen LT, et al. PLoS One. 2007 Sep 12;2(9):e858. doi: 10.1371/journal.pone.0000858. PLoS One. 2007. PMID: 17848989 Free PMC article.
• Structure of human Rad51 protein filament from molecular modeling and site-specific linear dichroism spectroscopy.
  Reymer A, Frykholm K, Morimatsu K, Takahashi M, Nordén B. Reymer A, et al. Proc Natl Acad Sci U S A. 2009 Aug 11;106(32):13248-53. doi: 10.1073/pnas.0902723106. Epub 2009 Jul 8. Proc Natl Acad Sci U S A. 2009. PMID: 19587234 Free PMC article.
• The Lon protease-like domain in the bacterial RecA paralog RadA is required for DNA binding and repair.
  Inoue M, Fukui K, Fujii Y, Nakagawa N, Yano T, Kuramitsu S, Masui R. Inoue M, et al. J Biol Chem. 2017 Jun 9;292(23):9801-9814. doi: 10.1074/jbc.M116.770180. Epub 2017 Apr 21. J Biol Chem. 2017. PMID: 28432121 Free PMC article.

Publication types

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Elsevier Science

• Other Literature Sources

  • The Lens - Patent Citations Database

• Molecular Biology Databases

  • GlyGen glycoinformatics resource
  • Saccharomyces Genome Database

• Research Materials

  • NCI CPTC Antibody Characterization Program