Qiu QS and Fratti RA (2010) The Na+/H+ exchanger Nhx1p regulates the initiation of Saccharomyces cerevisiae vacuole fusion. J Cell Sci 123(Pt 19):3266-75
Abstract: Nhx1p is a Na(+)(K(+))/H(+) antiporter localized at the vacuolar membrane of the yeast Saccharomyces cerevisiae. Nhx1p regulates the acidification of cytosol and vacuole lumen, and is involved in membrane traffic from late endosomes to the vacuole. Deletion of the gene leads to aberrant vacuolar morphology and defective vacuolar protein sorting. These phenotypes are hallmarks of malfunctioning vacuole homeostasis and indicate that membrane fusion is probably altered. Here, we investigated the role of Nhx1p in the regulation of homotypic vacuole fusion. Vacuoles isolated from nhx1Delta yeast showed attenuated fusion. Assays configured to differentiate between the first round of fusion and ongoing rounds showed that nhx1Delta vacuoles were only defective in the first round of fusion, suggesting that Nhx1p regulates an early step in the pathway. Although fusion was impaired on nhx1Delta vacuoles, SNARE complex formation was indistinguishable from wild-type vacuoles. Fusion could be rescued by adding the soluble SNARE Vam7p. However, Vam7p only activated the first round of nhx1Delta vacuole fusion. Once fusion was initiated, nhx1Delta vacuoles appeared behave in a wild-type manner. Complementation studies showed that ion transport function was required for Nhx1p-mediated support of fusion. In addition, the weak base chloroquine restored nhx1Delta fusion to wild-type levels. Together, these data indicate that Nhx1p regulates the initiation of fusion by controlling vacuole lumen pH.
|Status: Published||Type: Journal Article||PubMed ID: 20826459|
Topics addressed in this paper
Number of different genes curated to this paper: 4
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