Discrete nascent chain lengths are required for the insertion of presecretory proteins into microsomal membranes.
Ribosomes synthesizing nascent secretory proteins are targeted to the membrane by the signal recognition particle (SRP), a small ribonucleoprotein that binds to the signal peptide as it emerges from the ribosome. SRP arrests further elongation, causing ribosomes to stack behind the arrested ribosome. Upon interaction of SRP with its receptor on the ER membrane, the translation arrest is released and the ribosome becomes bound to the ER membrane. We have examined the distribution of unattached and membrane-bound ribosomes during the translation of mRNAs encoding two secretory proteins, bovine preprolactin and rat preproinsulin I. We find that the enhancement of ribosome stacking that occurs when SRP arrests translation of these proteins is relaxed in the presence of microsomal membranes. We also demonstrate that two previously described populations of membrane-associated ribosomes, distinguished by their sensitivity to high salt or EDTA extraction, correspond to ribosomes that have synthesized differing lengths of the nascent polypeptide. This analysis has revealed that nascent chain insertion into the membrane begins at distinct points for different presecretory proteins.
Department of Biochemistry and Biophysics, University of California Medical School, San Francisco, California 94143-0448.
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D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002417:Cattle; D007328:Insulin; D007425:Intracellular Membranes / Q000378:metabolism; D008861:Microsomes / Q000378:metabolism*; D008969:Molecular Sequence Data; D011384:Proinsulin / Q000096:biosynthesis*; D011388:Prolactin / Q000096:biosynthesis*; D014176:Protein Biosynthesis / Q000502:physiology*; D011498:Protein Precursors / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D012261:Ribonucleoproteins / Q000502:physiology; D012270:Ribosomes / Q000378:metabolism*; D018271:Signal Recognition Particle
D007328:Insulin; D011498:Protein Precursors; D012333:RNA, Messenger; D012261:Ribonucleoproteins; D018271:Signal Recognition Particle; C013737:preproinsulin; C017671:preprolactin; D011388:Prolactin; D011384:Proinsulin