Protoplasma

, Volume 250, Issue 6, pp 1283–1293

Organelle acidification is important for localisation of vacuolar proteins in Saccharomyces cerevisiae

Original Article

Abstract

The acidic environments in the vacuole and other acidic organelles are important for many cellular processes in eukaryotic cells. In this study, we comprehensively investigated the roles of organelle acidification in vacuolar protein localisation in Saccharomyces cerevisiae. After repressing the acidification of acidic compartments by treatment with concanamycin A, a specific inhibitor of vacuolar H+-ATPase (V-ATPase), we examined the localisation of GFP-fused proteins that were predicted to localise in the vacuolar lumen or on the vacuolar membrane. Of the 73 proteins examined, 19 changed their localisation to the cytoplasmic region. Localisation changes were evaluated quantitatively using the image processing programme CalMorph. The delocalised proteins included vacuolar hydrolases, V-ATPase subunits, transporters and enzymes for membrane biogenesis, as well as proteins required for protein transport. These results suggest that many alterations in the localisation of vacuolar proteins occur after loss of the acidification of acidic compartments.

Keywords

Vacuole Acidic compartments Concanamycin A CalMorph V-ATPase Saccharomyces cerevisiae 

Abbreviations

ACs

Acidic compartments

ALP

Alkaline phosphatase

CCA

Concanamycin A

CPS

Carboxypeptidase S

CPY

Carboxypeptidase Y

DIC

Differential interference contrast

DMSO

Dimethylsulfoxide

EUROSCARF

European Saccharomyces cerevisiae archive for functional analysis

GO

Gene ontology

EN

Endosomal network

PrA

Proteinase A

Quinacrine

6-Chloro-9(4-diethylamino-1-methylbutylamino)-2-methoxyacridine

SGD

Saccharomyces genome database

V-ATPases

Vacuolar proton-translocating ATPases

Supplementary material

709_2013_510_MOESM1_ESM.pdf (80 kb)
ESM 1(PDF 80.2 kb)
709_2013_510_MOESM2_ESM.pdf (162 kb)
Supplementary Fig. 1(PDF 162 kb)

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Copyright information

© Springer-Verlag Wien 2013

Authors and Affiliations

  • Risa Matsumoto
    • 1
  • Kuninori Suzuki
    • 2
  • Yoshikazu Ohya
    • 1
  1. 1.The Department of Integrated Biosciences, Graduate School of Frontier SciencesUniversity of TokyoKashiwaJapan
  2. 2.Bioimaging Center, Graduate School of Frontier SciencesUniversity of TokyoKashiwaJapan

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