HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 280, Issue 40, 33826-33838, October 7, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/40/33826    most recent M502477200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Barker, S. Articles by Murray, D. Search for Related Content PubMed PubMed Citation Articles by Barker, S. Articles by Murray, D. Social Bookmarking                      What's this?

Identification of Mammalian Proteins Cross-linked to DNA by Ionizing Radiation^*

Sharon Barker, Michael Weinfeld1, Jing Zheng, Liang Li, and David Murray2

From the Department of Oncology, Cross Cancer Institute, Edmonton, Alberta T6G 1Z2 and the Department of Chemistry W3–39, University of Alberta, Edmonton, Alberta T6G 2G2, Canada

Ionizing radiation (IR) is an important environmental risk factor for various cancers and also a major therapeutic agent for cancer treatment. Exposure of mammalian cells to IR induces several types of damage to DNA, including double- and single-strand breaks, base and sugar damage, as well as DNA-DNA and DNA-protein cross-links (DPCs). Little is known regarding the biological consequences of DPCs. Identifying the proteins that become cross-linked to DNA by IR would be an important first step in this regard. We have therefore undertaken a proteomics study to isolate and identify proteins involved in IR-induced DPCs. DPCs were induced in AA8 Chinese hamster ovary or GM00637 human fibroblast cells using 0–4 gray of -rays under either aerated or hypoxic conditions. DPCs were isolated using a recently developed method, and proteins were identified by mass spectrometry. We identified 29 proteins as being cross-linked to DNA by IR under aerated and/or hypoxic conditions. The identified proteins include structural proteins, actin-associated proteins, transcription regulators, RNA-splicing components, stress-response proteins, cell cycle regulatory proteins, and GDP/GTP-binding proteins. The involvement of several proteins (actin, histone H2B, and others) in DPCs was confirmed by using Western blot analysis. The dose responsiveness of DPC induction was examined by staining one-dimensional SDS-polyacrylamide gels with SYPRO Tangerine followed by analysis using fluorescence imaging. Quantitation of the fluorescence signal indicated no significant difference in total yields of IR-induced DPCs generated under aerated or hypoxic conditions, although differences were observed for several individual protein bands.

Received for publication, March 7, 2005 , and in revised form, August 1, 2005.

^* This work was supported by the National Cancer Institute of Canada, with funds from the Canadian Cancer Society, Grant 013104 (to M. W.), Terry Fox Research Grant 8067 (to D. M.), the Alberta Cancer Board through Pilot Project Grants R-294 and R-465 (to D. M.), and New Initiatives award (to L. L.) and studentship support (to S. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence may be addressed: Division of Experimental Oncology, Cross Cancer Institute, 11560 University Ave., Edmonton, Alberta T6G 1Z2, Canada. Tel.: 780-432-8427; Fax: 780-432-8428; E-mail: michaelw{at}cancerboard.ab.ca. ² To whom correspondence may be addressed. Tel.: 780-432-8427; Fax: 780-432-8428; E-mail: davem{at}cancerboard.ab.ca.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				D. J. Baker, G. Wuenschell, L. Xia, J. Termini, S. E. Bates, A. D. Riggs, and T. R. O'Connor Nucleotide Excision Repair Eliminates Unique DNA-Protein Cross-links from Mammalian Cells J. Biol. Chem., August 3, 2007; 282(31): 22592 - 22604. [Abstract] [Full Text] [PDF]