Document Detail


Base excision repair proteins are required for integrin-mediated suppression of bleomycin-induced DNA breakage in murine lung endothelial cells.
MedLine Citation:
PMID:  17202402     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Engagement of integrin cell adhesion receptors suppresses bleomycin (BLM)-induced DNA strand breakage in endothelial cells. Previous investigation of cells from poly(ADP-ribose) polymerase (PARP)-1 knockout mice and with an inhibitor of the enzyme indicated that this facilitator of base excision repair (BER) is required for integrin-mediated suppression of DNA strand breakage. Here, small inhibitory RNA (siRNA) was used to assess the requirement for the BER proteins, DNA ligase III (Lig3) alpha, PARP-1, and X-ray repair complementing defective repair in Chinese hamster cells 1 (XRCC1), and for the long-patch BER ligase, DNA ligase I (Lig1), in integrin-mediated protection from BLM-induced DNA breakage. Murine lung endothelial cells (MLECs) were transfected with siRNA, treated with anti-beta1 integrin antibody, and then BLM. 3'-OH in DNA and accumulation of phosphorylated histone H2AX (gammaH2AX), which reflects double-strand breakage, were measured. Integrin antibody inhibited the increases in 3'-OH caused by BLM in MLECs transfected with either control or Lig1 siRNA. However, after knockdown of Lig3alpha, PARP-1, or XRCC1, suppression of DNA breakage by integrin antibody was limited. BLM increased gammaH2AX levels, and integrin treatment inhibited this by 57 to 73% in MLECs transfected with control siRNA. Integrin engagement also inhibited increases in gammaH2AX in BLM-treated cells transfected with Lig1 siRNA. In contrast, Lig3alpha, PARP-1, and XRCC1 siRNAs prevented integrin-mediated inhibition of BLM-induced gammaH2AX levels. The results suggest that the BER proteins, Lig3alpha, PARP-1, and XRCC1, are required for integrin-mediated suppression of BLM-induced DNA breakage.
Authors:
Jane L Rose; Kevin C Reeves; Rostislav I Likhotvorik; Dale G Hoyt
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2007-01-03
Journal Detail:
Title:  The Journal of pharmacology and experimental therapeutics     Volume:  321     ISSN:  0022-3565     ISO Abbreviation:  J. Pharmacol. Exp. Ther.     Publication Date:  2007 Apr 
Date Detail:
Created Date:  2007-03-19     Completed Date:  2007-05-14     Revised Date:  2007-12-03    
Medline Journal Info:
Nlm Unique ID:  0376362     Medline TA:  J Pharmacol Exp Ther     Country:  United States    
Other Details:
Languages:  eng     Pagination:  318-26     Citation Subset:  IM    
Affiliation:
Division of Pharmacology, Ohio State University College of Pharmacy, 500 West Twelfth Avenue, Columbus, OH 43210, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antimetabolites, Antineoplastic / toxicity*
Bleomycin / antagonists & inhibitors*,  toxicity*
Blotting, Western
Cell Survival / drug effects
DNA Damage*
DNA Ligases / genetics
DNA Repair / drug effects*
DNA-Binding Proteins / genetics
Endothelial Cells / cytology,  drug effects,  metabolism*
Fluorescent Antibody Technique
Green Fluorescent Proteins / biosynthesis,  genetics
Histones / genetics
In Situ Nick-End Labeling
Integrins / physiology*
Lung / cytology,  drug effects,  metabolism*
Mice
Poly(ADP-ribose) Polymerases / antagonists & inhibitors,  genetics
RNA, Small Interfering / pharmacology
Reverse Transcriptase Polymerase Chain Reaction
Grant Support
ID/Acronym/Agency:
HL68054/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Antimetabolites, Antineoplastic; 0/DNA-Binding Proteins; 0/Histones; 0/Integrins; 0/RNA, Small Interfering; 0/X-ray repair cross complementing protein 1; 0/gamma-H2AX protein, mouse; 11056-06-7/Bleomycin; 147336-22-9/Green Fluorescent Proteins; EC 2.4.2.30/Poly(ADP-ribose) Polymerases; EC 2.4.2.30/poly(ADP-ribose)polymerase-1, mouse; EC 6.5.1.-/DNA Ligases; EC 6.5.1.1/DNA ligase (ATP)

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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