title: Inhibition of human HL-60 cell responses to chemotactic factors by antisense messenger RNA depletion of G proteins.
abstract: 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.
authors.affiliation: Department of Medicine, University of California Medical Center, San Francisco 94143-0711.
authors.forename: E J
authors.initials: EJ
authors.lastname: Goetzl
authors.forename: R S
authors.initials: RS
authors.lastname: Shames
authors.forename: J
authors.initials: J
authors.lastname: Yang
authors.forename: F W
authors.initials: FW
authors.lastname: Birke
authors.forename: Y F
authors.initials: YF
authors.lastname: Liu
authors.forename: P R
authors.initials: PR
authors.lastname: Albert
authors.forename: S
authors.initials: S
authors.lastname: An
mesh_terms: 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
chemical_list: D002630:Chemotactic Factors; D016372:RNA, Antisense; D007975:Leukotriene B4; D009240:N-Formylmethionine Leucyl-Phenylalanine; D000242:Cyclic AMP; D005966:Glucuronidase; D019204:GTP-Binding Proteins