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PMID: 17565997 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Cellular environment is important in controlling V-ATPase dissociation and its dependence on activity.

The Journal of biological chemistry ·Vol. 282 ·No. 34 ·2007-08-24 ·Pages 24743-51

Qi J, Forgac M

Abstract

One mechanism of regulating V-ATPase activity in vivo involves reversible dissociation into its component V(1) and V(0) domains, which in yeast occurs in response to glucose depletion. V-ATPase complexes containing the Vph1p isoform of subunit a (VCC) are targeted to the vacuole, and Stv1p-containing complexes (SCC) are targeted to the Golgi. Overexpression of Stv1p results in mistargeting of SCC to the vacuole. We have investigated the role of the a subunit isoform and cellular environment in controlling dissociation using vacuolar protein sorting (vps) mutants that accumulate proteins in either the prevacuolar compartment (PVC) (vps27Delta) or a post-Golgi compartment (PGC) (vps21Delta). Dissociation of both VCC and SCC depends upon cellular environment, with dissociation most complete in the vacuole and least complete in the PVC. The dependence of dissociation on V-ATPase activity was also investigated using both concanamycin and inactivating mutations. Concanamycin partly blocks dissociation of both VCC and SCC in all three compartments, with inhibition generally greater for SCC than VCC. The R735Q mutant of Vph1p results in loss of both ATPase and proton transport, whereas the R735K mutant lacks proton transport but has 10% of wild type ATPase activity. For VCC in the vacuole, dissociation is completely blocked for the R735Q but not the R735K mutant. Significant dissociation of VCC is observed for both mutants in the PVC and PGC, indicating that V-ATPase activity is not absolutely required for dissociation. Similar results were obtained for SCC, although dissociation of SCC is again generally more sensitive to activity than VCC. These results suggest that the cellular environment is important both in controlling in vivo dissociation of the V-ATPase and the dependence of this process on catalytic activity. Moreover, catalytic activity is not absolutely required for V-ATPase dissociation.

MeSH Terms
Adenosine Triphosphatases/metabolism Catalysis Enzyme Inhibitors/pharmacology Fungal Proteins/chemistry Glucose/metabolism Golgi Apparatus/metabolism Macrolides/pharmacology Models, Biological Mutagenesis, Site-Directed Mutation Protein Isoforms Saccharomyces cerevisiae/metabolism Vacuolar Proton-Translocating ATPases/chemistry Vacuoles
Chemicals
Enzyme Inhibitors Fungal Proteins Macrolides Protein Isoforms concanamycin A Adenosine Triphosphatases Vacuolar Proton-Translocating ATPases Glucose
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Qi Jie
Department of Physiology, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
Forgac Michael
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
DOI
10.1074/jbc.M700663200
Published
2007-08-24
Epub
2007-00-12
Pages
24743-51
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2394669
Subset
IM
Grants
NIGMS NIH HHS · R01 GM034478 · United States
NIDDK NIH HHS · P30 DK034928 · United States
NIDDK NIH HHS · DK 34928 · United States
NIGMS NIH HHS · R37 GM034478-23 · United States
NIGMS NIH HHS · R37 GM034478-20 · United States
NIGMS NIH HHS · GM 34478 · United States
NIGMS NIH HHS · R37 GM034478-22 · United States
NIGMS NIH HHS · R37 GM034478 · United States
NIGMS NIH HHS · R37 GM034478-21 · United States
External Links
PubMed source DOI: 10.1074/jbc.M700663200 PMC Full Text View journal details
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