Absence of DNA ligase IV protein in XR-1 cells: evidence for stabilization by XRCC4.
XR-1 is a CHO mutant cell line defective in double strand break repair and V(D)J recombination. These defects are due to a deletion of the XRCC4 gene which encodes a 38-kDa nuclear phosphoprotein. Recent studies have shown that XRCC4 interacts with and enhances the activity of DNA ligase IV in vitro. In this study we investigate the effect of the absence of XRCC4 on the level of DNA ligase IV in XR-1 cells. Western blot analysis indicates that levels of DNA ligase IV protein are almost undetectable in these cells, however, introduction of the XRCC4 cDNA into XR-1 resulted in a return to wild type levels of the protein. Furthermore, analysis of DNA ligase IV mRNA showed equivalent levels in both XR-1 and XRCC4 transfected XR-1 indicating that the altered level of DNA ligase IV is not due to a change in the expression of the gene. These data strongly suggest that an important function of XRCC4 is to stabilize the DNA ligase IV protein.
Bryans
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Lankenau Medical Research Center, Wynnewood, PA 19096, USA.
Valenzano
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MC
Stamato
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TD
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000378:metabolism*; D004260:DNA Repair / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004795:Enzyme Stability / Q000235:genetics*; D017353:Gene Deletion; D009687:Nuclear Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D011995:Recombination, Genetic / Q000235:genetics; D014162:Transfection / Q000235:genetics
D004268:DNA-Binding Proteins; C000607893:LIG4 protein, human; D009687:Nuclear Proteins; D012333:RNA, Messenger; C101467:XRCC4 protein, human; D011088:DNA Ligases; D000072481:DNA Ligase ATP