Detection of G-quadruplex DNA in mammalian cells
Alexander Henderson1, Yuliang Wu2, Yu Chuan Huang3, Elizabeth A. Chavez1, Jesse Platt4, F. Brad Johnson4, Robert M. Brosh Jr2, Dipankar Sen3 and Peter M. Lansdorp1,5,6,*
1Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, BC V5Z 1L3, Canada, 2Laboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, NIH Biomedical Research Center, Baltimore, MD 21224, USA, 3Department of Chemistry, Simon Fraser University, Burnaby, BC V5A 1S6, Canada, 4Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104-6100, USA, 5Division of Hematology, Department of Medicine, University of British Columbia, Vancouver, BC V6T 1Z4, Canada and 6European Research Institute for the Biology of Ageing, University of Groningen, University Medical Centre Groningen, A. Deusinglaan 1, NL-9713 AV Groningen, The Netherlands
↵*To whom correspondence should be addressed. Tel: +31 50 361 7300; Fax: +31 50 361 7310; Email: p.m.lansdorp{at}umcg.nl
Received August 30, 2013. Revision received September 25, 2013. Accepted September 30, 2013.
Abstract
It has been proposed that guanine-rich DNA forms four-stranded structures in vivo called G-quadruplexes or G4 DNA. G4 DNA has been implicated in several biological processes, but tools to study G4 DNA structures in cells are limited. Here we report the development of novel murine monoclonal antibodies specific for different G4 DNA structures. We show that one of these antibodies designated 1H6 exhibits strong nuclear staining in most human and murine cells. Staining intensity increased on treatment of cells with agents that stabilize G4 DNA and, strikingly, cells deficient in FANCJ, a G4 DNA-specific helicase, showed stronger nuclear staining than controls. Our data strongly support the existence of G4 DNA structures in mammalian cells and indicate that the abundance of such structures is increased in the absence of FANCJ. We conclude that monoclonal antibody 1H6 is a valuable tool for further studies on the role of G4 DNA in cell and molecular biology.
© The Author(s) 2013. Published by Oxford University Press.
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