Relationship between multiple biologic effects of rapamycin and the inhibition of pp70S6 protein kinase activity. Analysis in mutant clones of a T cell lymphoma.
Rapamycin (RAP) inhibits several biologic responses in the YAC-1 T cell lymphoma, including the serum-driven proliferation and cyclin A mRNA expression, the induction of Ly-6E Ag expression by IFN, and the induction of IFN-gamma production by IL-1. RAP also suppresses the enzymatic activity of the 70 kDa S6 protein kinase (pp70s6k). To define the mechanistic relationship between these multiple effects of RAP, we have generated stable somatic mutants with altered sensitivities to this drug. A first series of mutants, represented by the R19, 4R16, and 10R13 clones, showed markedly reduced sensitivity to the inhibitory effect of RAP on all biologic responses tested and on pp70s6k activity. Two other mutant types, R103 and R125, were both highly sensitive to RAP-mediated suppression of proliferation, of IL-1-induced IFN-gamma production, and of pp70s6k activity but differed in their Ly-6E response. This response was not affected by RAP in the R125 clone and was enhanced in the R103 clone. Therefore, the inhibitory effects of RAP on proliferation and IL-1-mediated IFN-gamma induction both appear associated with the inhibition of pp70s6k activity, whereas the modulation of Ly-6E induction is independent from the latter. Moreover, the cellular binding of [3H]dihydro-FK-506 was found to be blocked by RAP in all mutant types to the same extent as in wild-type YAC-1 cells, suggesting that the altered sensitivity to the effects of RAP in these mutants is not due to an inability of the drug to enter the cells or to interact with FKBP. Further biochemical characterization of the mutant cells described here is expected to help clarify the mechanisms of RAP action.
Dumont
F J
FJ
Department of Immunology, Merck Research Laboratories, Rahway, NJ 07065.
Altmeyer
A
A
Kastner
C
C
Fischer
P A
PA
Lemon
K P
KP
Chung
J
J
Blenis
J
J
Staruch
M J
MJ
D000818:Animals; D000950:Antigens, Ly / Q000378:metabolism; D002999:Clone Cells; D016213:Cyclins / Q000235:genetics; D015870:Gene Expression; D007371:Interferon-gamma / Q000096:biosynthesis / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects*; D016399:Lymphoma, T-Cell / Q000201:enzymology*; D051379:Mice; D011090:Polyenes / Q000737:chemistry / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins; D019893:Ribosomal Protein S6 Kinases; D020123:Sirolimus; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000950:Antigens, Ly; D016213:Cyclins; D007375:Interleukin-1; D011090:Polyenes; D012333:RNA, Messenger; D011994:Recombinant Proteins; D007371:Interferon-gamma; D017346:Protein Serine-Threonine Kinases; D019893:Ribosomal Protein S6 Kinases; D013755:Tetradecanoylphorbol Acetate; D020123:Sirolimus