An ATP transporter is required for protein translocation into the yeast endoplasmic reticulum

Peter Mayinger, David Meyer

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

The transfer of precursor proteins through the membrane of the rough endoplasmic reticulum (ER) in yeast is strictly dependent on the presence of ATP. Since Kar2p (the yeast homologue of mammalian BiP) is required for translocation, and is an ATP binding protein, an ATP transport system must be coupled to the translocation machinery of the ER. We report here the characterization of a transport system for ATP in vesicles derived from yeast ER. ATP uptake into vesicles was found to be saturable in the micromolar range with a Km of 1 × 10-5 M. ATP transport into ER vesicles was specifically inhibited by 4,4′-diisothiocyanatostilbene -2,2′-disulfonic acid (DIDS), a stilbene derivative known to inhibit a number of other anion transporters, and by 3′-O-(4-benzoyl)benzoyl-ATP (Bz2-ATP). Inhibition of ATP uptake into yeast microsomes by DIDS and Bz2-ATP blocked protein translocation in vitro measured co- as well as post-translationally. The inhibitory effect of DIDS on translocation was prevented by coincubation with ATP. Moreover, selective membrane permeabilization, allowing ATP access to the lumen, restored translocation activity to DIDS-treated membranes. These results demonstrate that translocation requires a DIDS and Bz2-ATP-sensitive component whose function is to transport ATP to the lumen of the ER. These Findings are consistent with current models of protein translocation in yeast which stipulate the participation of Karlp in the translocation process.

Original languageEnglish (US)
Pages (from-to)659-666
Number of pages8
JournalEMBO Journal
Volume12
Issue number2
StatePublished - 1993
Externally publishedYes

Fingerprint

Protein Transport
Endoplasmic Reticulum
Yeast
Adenosine Triphosphate
Yeasts
Proteins
Acids
Membranes
Stilbenes
Protein Precursors
Rough Endoplasmic Reticulum
Microsomes
Machinery
Anions
Carrier Proteins
Membrane Proteins

Keywords

  • ATP transport
  • Endoplasmic reticulum
  • Protein translocation
  • Saccharomyces cerevisiae

ASJC Scopus subject areas

  • Cell Biology
  • Genetics

Cite this

An ATP transporter is required for protein translocation into the yeast endoplasmic reticulum. / Mayinger, Peter; Meyer, David.

In: EMBO Journal, Vol. 12, No. 2, 1993, p. 659-666.

Research output: Contribution to journalArticle

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