Inhibition of human HL-60 cell responses to chemotactic factors by antisense messenger RNA depletion of G proteins.
Chemotactic factors bound to receptors of the seven-transmembrane domain family signal leukocytes through associated guanine nucleotide-binding (G) proteins. Human leukocytes of the HL-60 line, which express G protein-coupled receptors for leukotriene B4 (LTB4) and N-formyl-methionyl-leucyl-phenylalanine (fMLP) after differentiation with vitamin D3 and transforming growth factor-beta, were transfected with expression plasmids containing antisense-oriented cDNAs encoding the alpha-chains of Go, Gi1, Gi2, and Gi3. Antisense mRNA for Go and Gi2 alpha-chains suppressed by over 80% the level of the respective G protein. Go-deficient HL-60 cells had depressed functional and intracellular calcium responses to LTB4 and fMLP, but no alterations in the responses of cyclic adenosine 3',5'-monophosphate (cAMP). In contrast, HL-60 cells deficient in Gi2 lost only responses of the intracellular concentration of cAMP. Antisense mRNA suppression of distinct G proteins thus may delineate some transductional requirements for cellular responses.
Goetzl
E J
EJ
Department of Medicine, University of California Medical Center, San Francisco 94143-0711.
Shames
R S
RS
Yang
J
J
Birke
F W
FW
Liu
Y F
YF
Albert
P R
PR
An
S
S
D002460:Cell Line; D002630:Chemotactic Factors / Q000502:physiology*; D002634:Chemotaxis, Leukocyte; D000242:Cyclic AMP / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D005966:Glucuronidase / Q000378:metabolism; D006801:Humans; D007975:Leukotriene B4 / Q000494:pharmacology; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D016372:RNA, Antisense; D015398:Signal Transduction
D002630:Chemotactic Factors; D016372:RNA, Antisense; D007975:Leukotriene B4; D009240:N-Formylmethionine Leucyl-Phenylalanine; D000242:Cyclic AMP; D005966:Glucuronidase; D019204:GTP-Binding Proteins