Identification of a specific inhibitor for DNA ligase I in human cells.
A protein inhibitor for human DNA ligase I has recently been identified. It was copurified with a fraction of the enzymes from HeLa cells through several steps of chromatography. The inhibitor was first identified by the absence of ligation activity of the associated enzyme, while it retained the ability to form the ligase-[32P]AMP adducts. The inhibitor was eluted as a single peak at approximately 0.25-0.30 M NaCl from a Mono S column. It inhibited the ligation of both double-stranded and single-stranded breaks by purified DNA ligase I but not by T4 DNA ligase and DNA ligase II. Subsequent gel-filtration chromatography indicated that this inhibitor, with a molecular mass of 55-75 kDa, could form a complex with DNA ligase I and inhibited the DNA ligation activity. Rechromatography of the ligase I-inhibitor complex in high-salt conditions resulted in the dissociation of the complex and the restoration of enzyme activity, indicating that the physical interaction of inhibitor with DNA ligase I is one of the mechanisms of inhibition. These data indicate that this protein inhibitor for DNA ligase I may play a specific role in regulating DNA ligation during replication, repair, or recombination.
Yang
S W
SW
Department of Molecular Pathology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Becker
F F
FF
Chan
J Y
JY
D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D011485:Protein Binding
D004791:Enzyme Inhibitors; C000607892:LIG1 protein, human; D011088:DNA Ligases; D000072481:DNA Ligase ATP