Role of Vacuolar Acidification in Protein Sorting and Zymogen Activation: a Genetic Analysis of the Yeast Vacuolar Proton-translocating ATPase

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 1990 Jul 1

PMID 2141385

Citations 63

Authors

C T Yamashiro

P M Kane

D F Wolczyk

R A Preston

T H Stevens

Affiliations

Soon will be listed here.

Abstract

Vacuolar acidification has been proposed to play a key role in a number of cellular processes, including protein sorting, zymogen activation, and maintenance of intracellular pH. We investigated the significance of vacuolar acidification by cloning and mutagenizing the gene for the yeast vacuolar proton-translocating ATPase 60-kilodalton subunit (VAT2). Cells carrying a vat2 null allele were viable; however, these cells were severely defective for growth in medium buffered at neutral pH. Vacuoles isolated from cells bearing the vat2 null allele were completely devoid of vacuolar ATPase activity. The pH of the vacuolar lumen of cells bearing the vat2 mutation was 7.1, compared with the wild-type pH of 6.1, as determined by a flow cytometric pH assay. These results indicate that the vacuolar proton-translocating ATPase complex is essential for vacuolar acidification and that the low-pH state of the vacuole is crucial for normal growth. The vacuolar acidification-defective vat2 mutant exhibited normal zymogen activation but displayed a minor defect in vacuolar protein sorting.

Citing Articles

The Toxoplasma plant-like vacuolar compartment (PLVAC).

Stasic A, Moreno S, Carruthers V, Dou Z J Eukaryot Microbiol. 2022; 69(6):e12951.

PMID: 36218001 PMC: 10576567. DOI: 10.1111/jeu.12951.

Mutations in V-ATPase in follicular lymphoma activate autophagic flux creating a targetable dependency.

Wang F, Yang Y, Klionsky D, Malek S Autophagy. 2022; 19(2):716-719.

PMID: 35482846 PMC: 9851240. DOI: 10.1080/15548627.2022.2071382.

ReporterSeq reveals genome-wide dynamic modulators of the heat shock response across diverse stressors.

Alford B, Tassoni-Tsuchida E, Khan D, Work J, Valiant G, Brandman O Elife. 2021; 10.

PMID: 34223816 PMC: 8257254. DOI: 10.7554/eLife.57376.

Sulforaphane alters the acidification of the yeast vacuole.

Wilcox A, Murphy M, Tucker D, Laprade D, Roussel B, Chin C Microb Cell. 2020; 7(5):129-138.

PMID: 32391394 PMC: 7199281. DOI: 10.15698/mic2020.05.716.

Functional complementation reveals that 9 of the 13 human V-ATPase subunits can functionally substitute for their yeast orthologs.

Abe M, Saito M, Tsukahara A, Shiokawa S, Ueno K, Shimamura H J Biol Chem. 2019; 294(20):8273-8285.

PMID: 30952699 PMC: 6527166. DOI: 10.1074/jbc.RA118.006192.

References

Jones E . The synthesis and function of proteases in Saccharomyces: genetic approaches. Annu Rev Genet. 1984; 18:233-70. DOI: 10.1146/annurev.ge.18.120184.001313. View

James M, Sielecki A . Molecular structure of an aspartic proteinase zymogen, porcine pepsinogen, at 1.8 A resolution. Nature. 1986; 319(6048):33-8. DOI: 10.1038/319033a0. View

Stevens T, Rothman J, Payne G, Schekman R . Gene dosage-dependent secretion of yeast vacuolar carboxypeptidase Y. J Cell Biol. 1986; 102(5):1551-7. PMC: 2114202. DOI: 10.1083/jcb.102.5.1551. View

Rothman J, Hunter C, Valls L, Stevens T . Overproduction-induced mislocalization of a yeast vacuolar protein allows isolation of its structural gene. Proc Natl Acad Sci U S A. 1986; 83(10):3248-52. PMC: 323490. DOI: 10.1073/pnas.83.10.3248. View

Sharp P, Tuohy T, Mosurski K . Codon usage in yeast: cluster analysis clearly differentiates highly and lowly expressed genes. Nucleic Acids Res. 1986; 14(13):5125-43. PMC: 311530. DOI: 10.1093/nar/14.13.5125. View

von Figura K, Hasilik A . Lysosomal enzymes and their receptors. Annu Rev Biochem. 1986; 55:167-93. DOI: 10.1146/annurev.bi.55.070186.001123. View

Mellman I, Fuchs R, Helenius A . Acidification of the endocytic and exocytic pathways. Annu Rev Biochem. 1986; 55:663-700. DOI: 10.1146/annurev.bi.55.070186.003311. View

Timchak L, Kruse F, Marnell M, Draper R . A thermosensitive lesion in a Chinese hamster cell mutant causing differential effects on the acidification of endosomes and lysosomes. J Biol Chem. 1986; 261(30):14154-9. View

Klionsky D, Emr S . Membrane protein sorting: biosynthesis, transport and processing of yeast vacuolar alkaline phosphatase. EMBO J. 1989; 8(8):2241-50. PMC: 401154. DOI: 10.1002/j.1460-2075.1989.tb08348.x. View

10.

Rothman J, Yamashiro C, Kane P, Stevens T . Protein targeting to the yeast vacuole. Trends Biochem Sci. 1989; 14(8):347-50. DOI: 10.1016/0968-0004(89)90170-9. View

11.

Kane P, Yamashiro C, Stevens T . Biochemical characterization of the yeast vacuolar H(+)-ATPase. J Biol Chem. 1989; 264(32):19236-44. View

12.

Kornfeld S, Mellman I . The biogenesis of lysosomes. Annu Rev Cell Biol. 1989; 5:483-525. DOI: 10.1146/annurev.cb.05.110189.002411. View

13.

Kane P, Yamashiro C, Rothman J, Stevens T . Protein sorting in yeast: the role of the vacuolar proton-translocating ATPase. J Cell Sci Suppl. 1989; 11:161-78. DOI: 10.1242/jcs.1989.supplement_11.13. View

14.

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

15.

Rothman J, Yamashiro C, Raymond C, Kane P, Stevens T . Acidification of the lysosome-like vacuole and the vacuolar H+-ATPase are deficient in two yeast mutants that fail to sort vacuolar proteins. J Cell Biol. 1989; 109(1):93-100. PMC: 2115461. DOI: 10.1083/jcb.109.1.93. View

16.

Forgac M . Structure and function of vacuolar class of ATP-driven proton pumps. Physiol Rev. 1989; 69(3):765-96. DOI: 10.1152/physrev.1989.69.3.765. View

17.

Bustin M . Intramolecular activation of porcine pepsinogen. J Biol Chem. 1971; 246(3):615-20. View

18.

SANGER F, Nicklen S, Coulson A . DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977; 74(12):5463-7. PMC: 431765. DOI: 10.1073/pnas.74.12.5463. View

19.

Ohkuma S, Poole B . Fluorescence probe measurement of the intralysosomal pH in living cells and the perturbation of pH by various agents. Proc Natl Acad Sci U S A. 1978; 75(7):3327-31. PMC: 392768. DOI: 10.1073/pnas.75.7.3327. View

20.

Tang J . Evolution in the structure and function of carboxyl proteases. Mol Cell Biochem. 1979; 26(2):93-109. DOI: 10.1007/BF00232887. View