Analyzing the Vacuolar Membrane (Tonoplast) Proteome

2017 ◽  
pp. 107-116 ◽  
Author(s):  
Miwa Ohnishi ◽  
Katsuhisa Yoshida ◽  
Tetsuro Mimura
Keyword(s):  
Vacuolar Membrane

scholarly journals VMA12 is essential for assembly of the vacuolar H(+)-ATPase subunits onto the vacuolar membrane in Saccharomyces cerevisiae.

1993 ◽  
Vol 268 (2) ◽  
pp. 961-967
Author(s):  
R. Hirata ◽  
N. Umemoto ◽  
M.N. Ho ◽  
Y. Ohya ◽  
T.H. Stevens ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Vacuolar Membrane ◽  
Atpase Subunits

scholarly journals Membrane recruitment of Atg8 by Hfl1 facilitates turnover of vacuolar membrane proteins in yeast cells approaching stationary phase

BMC Biology ◽  
2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Cheng-Wen He ◽  
Xue-Fei Cui ◽  
Shao-Jie Ma ◽  
Qin Xu ◽  
Yan-Peng Ran ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Membrane Protein ◽  
Stationary Phase ◽  
Molecular Mechanisms ◽  
Multivesicular Body ◽  
Degradation Process ◽  
Vacuolar Membrane ◽  
Yeast Cells ◽  
Membrane Structures ◽  
Membrane Recruitment

Abstract Background The vacuole/lysosome is the final destination of autophagic pathways, but can also itself be degraded in whole or in part by selective macroautophagic or microautophagic processes. Diverse molecular mechanisms are involved in these processes, the characterization of which has lagged behind those of ATG-dependent macroautophagy and ESCRT-dependent endosomal multivesicular body pathways. Results Here we show that as yeast cells gradually exhaust available nutrients and approach stationary phase, multiple vacuolar integral membrane proteins with unrelated functions are degraded in the vacuolar lumen. This degradation depends on the ESCRT machinery, but does not strictly require ubiquitination of cargos or trafficking of cargos out of the vacuole. It is also temporally and mechanistically distinct from NPC-dependent microlipophagy. The turnover is facilitated by Atg8, an exception among autophagy proteins, and an Atg8-interacting vacuolar membrane protein, Hfl1. Lack of Atg8 or Hfl1 led to the accumulation of enlarged lumenal membrane structures in the vacuole. We further show that a key function of Hfl1 is the membrane recruitment of Atg8. In the presence of Hfl1, lipidation of Atg8 is not required for efficient cargo turnover. The need for Hfl1 can be partially bypassed by blocking Atg8 delipidation. Conclusions Our data reveal a vacuolar membrane protein degradation process with a unique dependence on vacuole-associated Atg8 downstream of ESCRTs, and we identify a specific role of Hfl1, a protein conserved from yeast to plants and animals, in membrane targeting of Atg8.

Vacuole Partitioning During Meiotic Division in Yeast

Genetics ◽  
1996 ◽  
Vol 144 (2) ◽  
pp. 445-458 ◽  
Author(s):  
Amy D Roeder ◽  
Janet M Shaw
Keyword(s):  
Alkaline Phosphatase ◽  
Meiotic Division ◽  
Cell Walls ◽  
Vacuolar Membrane ◽  
Carboxypeptidase Y ◽  
Fluorescent Markers ◽  
Immunofluorescence Studies ◽  
Membrane Bound ◽  
Subsequent Staining ◽  
Spore Cell

Abstract We have examined the partitioning of the yeast vacuole during meiotic division. In pulse-chase experiments, vacuoles labeled with the lumenal ade2 fluorophore or the membrane-specific dye FM 4-64 were not inherited by haploid spores. Instead, these fluorescent markers were excluded from spores and trapped between the spore cell walls and the ascus. Serial optical sections using a confocal microscope confirmed that spores did not inherit detectable amounts of fluorescently labeled vacuoles. Moreover, indirect immunofluorescence studies established that an endogenous vacuolar membrane protein, alkaline phosphatase, and a soluable vacuolar protease, carboxypeptidase Y, were also detected outside spores after meiotic division. Spores that did not inherit ade2- or FM 4-64-labeled vacuoles did generate an organelle that could be visualized by subsequent staining with vacuole-specific fluorophores. These data contrast with genetic evidence that a soluble vacuolar protease is inherited by spores. When the partitioning of both types of markers was examined in sporulating cultures, the vacuolar protease activity was inherited by spores while fluorescently labeled vacuoles were largely excluded from spores. Our results indicate that the majority of the diploid vacuole, both soluble contents and membrane-bound components, are excluded from spores formed during meiotic division.

The tetraspanin TSP3 of Neurospora crassa is a vacuolar membrane protein and shares characteristics with IDI proteins

Mycologia ◽  
2016 ◽  
Vol 108 (3) ◽  
pp. 581-589
Author(s):  
Daniela Heine ◽  
Linda Petereit ◽  
Marcel R. Schumann ◽  
Diana Patzelt ◽  
Leila Rachid ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Neurospora Crassa ◽  
Vacuolar Membrane
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