Stimulation of protein synthesis by internalized insulin.
Previous studies showed that microinjected insulin stimulates transcription and translation in Stage IV Xenopus oocytes by acting at nuclear and cytoplasmic sites (Miller, D.S., 1988, 1989). The present report is concerned with the question of whether hormone, internalized from an external medium, can act on those sites to alter cell function. Both intracellular accumulation of undegraded 125I-insulin and insulin-stimulated 35S-methionine incorporation into oocyte protein were measured. Anti-insulin antiserum and purified anti-insulin antibody were microinjected into the cytoplasm of insulin-exposed cells to determine if insulin derived from the medium acted through internal sites. In cells exposed for 2 h to 7 or 70 nM external insulin, methionine incorporation was stimulated, but intracellular hormone accumulation was minimal and microinjected antibody was without effect. In cells exposed for 24 h, methionine incorporation again increased, but now accumulation of undegraded, intracellular hormone was substantial (2.6 and 25.3 fmol with 7 and 70 nM, respectively), and microinjected anti-insulin antibody significantly reduced the insulin-stimulated component of incorporation; basal incorporation was not affected. For cells exposed to 70 nM insulin for 24 h, inhibition of the insulin-stimulated component was maximal at 39%. Thus under those conditions, about 40% of insulin's effects were mediated by the internal sites. Together, the data show that inhibition of insulin-stimulated protein synthesis by microinjected antibody was associated with the intracellular accumulation of insulin. They indicate that when oocytes are exposed to external insulin, hormone eventually gains access to intracellular sites of action and through these stimulates translation. Control of translation appears to be shared between the internal sites and the surface receptor.
Miller
D S
DS
Laboratory of Cellular and Molecular Pharmacology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709.
Sykes
D B
DB
D000818:Animals; D000906:Antibodies / Q000008:administration & dosage / Q000276:immunology; D004305:Dose-Response Relationship, Drug; D005260:Female; D007328:Insulin / Q000276:immunology / Q000494:pharmacology*; D007457:Iodine Radioisotopes; D008715:Methionine / Q000378:metabolism; D008845:Microinjections; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects; D011506:Proteins / Q000235:genetics; D013462:Sulfur Radioisotopes; D014158:Transcription, Genetic / Q000187:drug effects; D014982:Xenopus laevis
D000906:Antibodies; D007328:Insulin; D007457:Iodine Radioisotopes; D011506:Proteins; D013462:Sulfur Radioisotopes; D008715:Methionine