Dithiothreitol and the translocation of preprolactin across mammalian endoplasmic reticulum.
The translocation mode of preprolactin (pPL) across mammalian endoplasmic reticulum was reinvestigated in light of recent findings that nascent secretory polypeptides synthesized in the presence of a highly reducing environment could be translocated posttranslationally and independently of their attachment to the ribosome (Maher, P. A., and S. J. Singer, 1986, Proc. Natl. Acad. Sci. USA, 83:9001-9005). The effects of the reducing agent dithiothreitol (DTT) on pPL synthesis and translocation were studied in this respect. The translocation of pPL was shown to take place only cotranslationally. The apparent posttranslational translocation was due to ongoing chain synthesis irrespective of the presence of high concentrations of DTT. When synthesis was completely blocked, no translocation was observed in the presence or absence of DTT. The synthesis of pPL was retarded by DTT, while its percent translocation was enhanced. The retardation in synthesis was reflected in reduced rates of initiation and elongation. As a consequence of this retardation, which increases the ratio of microsomes to nascent chains, and of possible effects on the conformation of nascent pPL and components of the translocation apparatus, DTT may expand the time and chain length windows for nascent chain translocation competence.
Ibrahimi
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European Molecular Biology Laboratory, Heidelberg, Federal Republic of Germany.
D001692:Biological Transport / Q000187:drug effects; D004229:Dithiothreitol / Q000494:pharmacology*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000187:drug effects; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects; D011388:Prolactin / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000187:drug effects; D011498:Protein Precursors / Q000096:biosynthesis / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000187:drug effects
D011498:Protein Precursors; C017671:preprolactin; D011388:Prolactin; D004229:Dithiothreitol