Rapamycin selectively represses translation of the "polypyrimidine tract" mRNA family.
The immunosuppressant rapamycin blocks p70s6k/p85s6k activation and phosphorylation of 40S ribosomal protein S6 in Swiss 3T3 cells. The same net result is obtained when the macrolide is added 3 hr after serum stimulation. In stimulated cells p70s6k/p85s6k inactivation is achieved within minutes, whereas S6 dephosphorylation requires 1-2 hr, supporting the concept that S6 dephosphorylation results from kinase inactivation. In parallel, rapamycin treatment causes a small, but significant, reduction in the initiation rate of protein synthesis, as measured both by [35S]methionine incorporation into protein and by recruitment of 80S ribosomes into polysomes. More striking, analysis of individual mRNA transcripts revealed that rapamycin selectively suppresses the translation of a family of mRNAs that is characterized by a polypyrimidine tract immediately after their N7-methylguanosine cap, a motif that can act as a translational modulator. This family includes transcripts for ribosomal proteins, elongation factors of protein synthesis, and proteins of as-yet-unknown function. The results imply that (i) 40S ribosomes containing phosphorylated S6 may selectively recognize this motif or proteins which bind to it and (ii) rapamycin may inhibit cell growth by blocking S6 phosphorylation and, thus, translation of these mRNAs.
Jefferies
H B
HB
Friedrich Miescher Institute, Basel, Switzerland.
Reinhard
C
C
Kozma
S C
SC
Thomas
G
G
D016475:3T3 Cells; D000199:Actins / Q000096:biosynthesis*; D000818:Animals; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000096:biosynthesis*; D010766:Phosphorylation; D011090:Polyenes / Q000494:pharmacology*; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects*; D011743:Pyrimidines; D012333:RNA, Messenger / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins / Q000187:drug effects / Q000378:metabolism; D020123:Sirolimus; D014158:Transcription, Genetic
D000199:Actins; D007166:Immunosuppressive Agents; D015345:Oligonucleotide Probes; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D011090:Polyenes; D011743:Pyrimidines; D012333:RNA, Messenger; D038601:Ribosomal Protein S6; D012269:Ribosomal Proteins; D008715:Methionine; D020123:Sirolimus