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. 1986 May;5(5):1031–1036. doi: 10.1002/j.1460-2075.1986.tb04318.x

Secretion in yeast: translocation and glycosylation of prepro-α-factor in vitro can occur via an ATP-dependent post-translational mechanism

Jonathan A Rothblatt 1, David I Meyer 1
PMCID: PMC1166897  PMID: 15957217

Abstract

In an in vitro system comprising a yeast cell-free translation system, yeast microsomes and mRNA encoding prepro-α-factor, the translocation of this protein across the membrane of the microsomal vesicle and its glycosylation could be uncoupled from its translation. Such post-translational processing is dependent upon the presence of ATP in the system. It is not, however, affected by a variety of uncouplers, ionophores or inhibitors, including carbonyl cyanide m-chlorophenyl hydrazone (CCCP), valinomycin, nigericin, dinitrophenol (DNP), potassium cyanide (KCN) or N-ethyl maleimide (NEM). This mechanism of translocation is significant as it indicates that a protein of 18 000 daltons is capable of crossing an endoplasmic reticulum-derived membrane post-translationally. For the moment, this phenomenon seems to be restricted to prepro-α-factor in the yeast in vitro system. Neither invertase nor IgG ϰ light chain could be translocated post-translationally in yeast, nor was such processing observed for prepro-α-factor in a wheat germ system supplemented with canine pancreatic microsomes.

Keywords: secretion, protein translocation, endoplasmic reticulum, membrane biogenesis, Saccharomyces cerevisiae

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