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

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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.

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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

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Acknowledgements

We thank Shinsuke Ohnuki for development of CalMorphGFP (version 1.1) and construction of the database, and members of the Laboratory of Signal Transduction for fruitful discussions. This work was supported by the Hamaguchi Foundation for the Advancement of Biochemistry (K.S.), the NOVARTIS Foundation (Japan) for the Promotion of Science (K.S.) and Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan (24121707 and 24657083 to K.S.; 21310127 and 24370002 to Y.O.).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. The Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Bldg. FSB-101, 5-1-5 Kashiwanoha, Kashiwa, Chiba, 277-8562, Japan

    Risa Matsumoto & Yoshikazu Ohya

  2. Bioimaging Center, Graduate School of Frontier Sciences, University of Tokyo, Bldg. FSB-101, 5-1-5 Kashiwanoha, Kashiwa, Chiba, 277-8562, Japan

    Kuninori Suzuki

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  1. Risa Matsumoto
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  2. Kuninori Suzuki
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  3. Yoshikazu Ohya
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Correspondence to Yoshikazu Ohya.

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Matsumoto, R., Suzuki, K. & Ohya, Y. Organelle acidification is important for localisation of vacuolar proteins in Saccharomyces cerevisiae . Protoplasma 250, 1283–1293 (2013). https://doi.org/10.1007/s00709-013-0510-2

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  • DOI: https://doi.org/10.1007/s00709-013-0510-2

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