Transient activation of topoisomerase I in leukotriene D4 signal transduction in human cells.
U937 human monoblast cells incubated with leukotriene D4 (LTD4) rapidly released arachidonic acid metabolites into the culture medium. Release was suppressed by the high-affinity LTD4 receptor antagonist SK&F 104353. Arachidonic acid release induced by LTD4 has been linked to a rapid induction of gene expression, and the propagation of the receptor binding signal is probably associated with enzymes that regulate gene expression. We have studied the participation of DNA topoisomerase I in LTD4 signal transduction. LTD4-specific release of arachidonic acid metabolites was inhibited (60-80%) by the topoisomerase I inhibitor camptothecin. LTD4 increased protein-linked DNA strand breakage induced by camptothecin in U937 cells; this enhancement was prevented by coincubation of the cells with LTD4 plus the receptor antagonist SK&F 104353. In addition, LTD4 produced a rapid transient increase in extractable topoisomerase I activity, which was maximum within the first 10 min after addition of LTD4 to the culture medium. Incubation of cultures for greater than 10 min with LTD4 before the addition of camptothecin resulted in no enhancement of camptothecin-induced DNA strand breakage, consistent with a reversal of topoisomerase I activation. Staurosporine, an inhibitor of protein kinase C, blocked LTD4-induced arachidonic acid release and attenuated the effect of LTD4 on camptothecin-induced DNA strand breakage. These results are consistent with the view that the regulation of topoisomerase I activity is involved in the propagation of LTD4-mediated signals in U937 cells.
Mattern
M R
MR
SmithKline & French Laboratories, King of Prussia, PA 19406-0939.
Mong
S
S
Mong
S M
SM
Bartus
J O
JO
Sarau
H M
HM
Clark
M A
MA
Foley
J J
JJ
Crooke
S T
ST
D000470:Alkaloids / Q000494:pharmacology; D001095:Arachidonic Acids / Q000037:antagonists & inhibitors / Q000378:metabolism; D002166:Camptothecin / Q000633:toxicity; D002478:Cells, Cultured; D004249:DNA Damage; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D003998:Dicarboxylic Acids / Q000494:pharmacology; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D013189:SRS-A / Q000378:metabolism*; D015398:Signal Transduction; D019311:Staurosporine
D000470:Alkaloids; D001095:Arachidonic Acids; D003998:Dicarboxylic Acids; D013189:SRS-A; D004264:DNA Topoisomerases, Type I; D019311:Staurosporine; C052393:pobilukast; D002166:Camptothecin