Sac1p mediates the adenosine triphosphate transport into yeast endoplasmic reticulum that is required for protein translocation

Peter Mayinger, Vytas A. Bankaitis, David I. Meyer

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Protein translocation into the yeast endoplasmic reticulum requires the transport of ATP into the lumen of this organelle. Microsomal ATP transport activity was reconstituted into proteoliposomes to characterize and identify the transporter protein. A polypeptide was purified whose partial amino acid sequence demonstrated its identity to the product of the SAC1 gene. Accordingly, microsomal membranes isolated from strains harboring a deletion in the SAC1 gene (sac1Δ) were found to be deficient in ATP-transporting activity as well as severely compromised in their ability to translocate nascent prepro-α-factor and preprocarboxypeptidase Y. Proteins isolated from the microsomal membranes of a sac1Δ strain were incapable of stimulating ATP transport when reconstituted into the in vitro assay system. When immunopurified to homogeneity and incorporated into artificial lipid vesicles, Sac1p was shown to reconstitute ATP transport activity. Consistent with the requirement for ATP in the lumen of the ER to achieve the correct folding of secretory proteins, the sac1Δ strain was shown to have a severe defect in transport of procarboxypeptidase Y out of the ER and into the Golgi complex in vivo. The collective data indicate an intimate role for Sac1p in the transport of ATP into the ER lumen.

Original languageEnglish (US)
Pages (from-to)1377-1386
Number of pages10
JournalJournal of Cell Biology
Volume131
Issue number6 I
DOIs
StatePublished - Dec 1995
Externally publishedYes

Fingerprint

Protein Transport
Endoplasmic Reticulum
Adenosine Triphosphate
Yeasts
Membranes
Protein Folding
Golgi Apparatus
Organelles
Genes
Amino Acid Sequence
Proteins
Lipids
Peptides

ASJC Scopus subject areas

  • Cell Biology

Cite this

Sac1p mediates the adenosine triphosphate transport into yeast endoplasmic reticulum that is required for protein translocation. / Mayinger, Peter; Bankaitis, Vytas A.; Meyer, David I.

In: Journal of Cell Biology, Vol. 131, No. 6 I, 12.1995, p. 1377-1386.

Research output: Contribution to journalArticle

@article{ceec679fb4e845f5872bc67b86bbab6e,
title = "Sac1p mediates the adenosine triphosphate transport into yeast endoplasmic reticulum that is required for protein translocation",
abstract = "Protein translocation into the yeast endoplasmic reticulum requires the transport of ATP into the lumen of this organelle. Microsomal ATP transport activity was reconstituted into proteoliposomes to characterize and identify the transporter protein. A polypeptide was purified whose partial amino acid sequence demonstrated its identity to the product of the SAC1 gene. Accordingly, microsomal membranes isolated from strains harboring a deletion in the SAC1 gene (sac1Δ) were found to be deficient in ATP-transporting activity as well as severely compromised in their ability to translocate nascent prepro-α-factor and preprocarboxypeptidase Y. Proteins isolated from the microsomal membranes of a sac1Δ strain were incapable of stimulating ATP transport when reconstituted into the in vitro assay system. When immunopurified to homogeneity and incorporated into artificial lipid vesicles, Sac1p was shown to reconstitute ATP transport activity. Consistent with the requirement for ATP in the lumen of the ER to achieve the correct folding of secretory proteins, the sac1Δ strain was shown to have a severe defect in transport of procarboxypeptidase Y out of the ER and into the Golgi complex in vivo. The collective data indicate an intimate role for Sac1p in the transport of ATP into the ER lumen.",
author = "Peter Mayinger and Bankaitis, {Vytas A.} and Meyer, {David I.}",
year = "1995",
month = "12",
doi = "10.1083/jcb.131.6.1377",
language = "English (US)",
volume = "131",
pages = "1377--1386",
journal = "Journal of Cell Biology",
issn = "0021-9525",
publisher = "Rockefeller University Press",
number = "6 I",

}

TY - JOUR

T1 - Sac1p mediates the adenosine triphosphate transport into yeast endoplasmic reticulum that is required for protein translocation

AU - Mayinger, Peter

AU - Bankaitis, Vytas A.

AU - Meyer, David I.

PY - 1995/12

Y1 - 1995/12

N2 - Protein translocation into the yeast endoplasmic reticulum requires the transport of ATP into the lumen of this organelle. Microsomal ATP transport activity was reconstituted into proteoliposomes to characterize and identify the transporter protein. A polypeptide was purified whose partial amino acid sequence demonstrated its identity to the product of the SAC1 gene. Accordingly, microsomal membranes isolated from strains harboring a deletion in the SAC1 gene (sac1Δ) were found to be deficient in ATP-transporting activity as well as severely compromised in their ability to translocate nascent prepro-α-factor and preprocarboxypeptidase Y. Proteins isolated from the microsomal membranes of a sac1Δ strain were incapable of stimulating ATP transport when reconstituted into the in vitro assay system. When immunopurified to homogeneity and incorporated into artificial lipid vesicles, Sac1p was shown to reconstitute ATP transport activity. Consistent with the requirement for ATP in the lumen of the ER to achieve the correct folding of secretory proteins, the sac1Δ strain was shown to have a severe defect in transport of procarboxypeptidase Y out of the ER and into the Golgi complex in vivo. The collective data indicate an intimate role for Sac1p in the transport of ATP into the ER lumen.

AB - Protein translocation into the yeast endoplasmic reticulum requires the transport of ATP into the lumen of this organelle. Microsomal ATP transport activity was reconstituted into proteoliposomes to characterize and identify the transporter protein. A polypeptide was purified whose partial amino acid sequence demonstrated its identity to the product of the SAC1 gene. Accordingly, microsomal membranes isolated from strains harboring a deletion in the SAC1 gene (sac1Δ) were found to be deficient in ATP-transporting activity as well as severely compromised in their ability to translocate nascent prepro-α-factor and preprocarboxypeptidase Y. Proteins isolated from the microsomal membranes of a sac1Δ strain were incapable of stimulating ATP transport when reconstituted into the in vitro assay system. When immunopurified to homogeneity and incorporated into artificial lipid vesicles, Sac1p was shown to reconstitute ATP transport activity. Consistent with the requirement for ATP in the lumen of the ER to achieve the correct folding of secretory proteins, the sac1Δ strain was shown to have a severe defect in transport of procarboxypeptidase Y out of the ER and into the Golgi complex in vivo. The collective data indicate an intimate role for Sac1p in the transport of ATP into the ER lumen.

UR - http://www.scopus.com/inward/record.url?scp=0029620936&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029620936&partnerID=8YFLogxK

U2 - 10.1083/jcb.131.6.1377

DO - 10.1083/jcb.131.6.1377

M3 - Article

VL - 131

SP - 1377

EP - 1386

JO - Journal of Cell Biology

JF - Journal of Cell Biology

SN - 0021-9525

IS - 6 I

ER -