{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T04:33:33Z","timestamp":1768451613139,"version":"3.49.0"},"reference-count":31,"publisher":"Rockefeller University Press","issue":"1","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1998,7,13]]},"abstract":"<jats:p>Three previously identified genes from Saccharomyces cerevisiae, VMA12, VMA21, and VMA22, encode proteins localized to the endoplasmic reticulum (ER). These three proteins are required for the biogenesis of a functional vacuolar ATPase (V-ATPase), but are not part of the final enzyme complex. Subcellular fractionation and chemical cross-linking studies have revealed that Vma12p and Vma22p form a stable membrane associated complex. Cross-linking analysis also revealed a direct physical interaction between the Vma12p\/Vma22p assembly complex and Vph1p, the 100-kD integral membrane subunit of the V-ATPase. The interaction of the Vma12p\/Vma22p complex with Vph1p was transient (half-life of \u223c5 min), reflecting trafficking of this V-ATPase subunit through the ER en route to the vacuolar membrane. Analysis of these protein\u2013protein interactions in ER-blocked sec12 mutant cells indicated that the Vph1p-Vma12p\/Vma22p interactions are quite stable when transport of the V-ATPase out of the ER is blocked. Fractionation of solubilized membrane proteins on a density gradient revealed comigration of Vma22p and Vma12p, indicating that they form a complex even in the absence of cross-linker. Vma12p and Vma22p migrated to fractions separate from Vma21p. Loss of Vph1p caused the Vma12p\/Vma22p complex to sediment to less dense fractions, consistent with association of Vma12p\/ Vma22p with nascent Vph1p in ER membranes. This is the first evidence for a dedicated assembly complex in the ER required for the assembly of an integral membrane protein complex (V-ATPase) as it is transported through the secretory pathway.<\/jats:p>","DOI":"10.1083\/jcb.142.1.39","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T16:47:50Z","timestamp":1027702070000},"page":"39-49","update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":91,"title":["Assembly of the Yeast Vacuolar H+-ATPase Occurs in the Endoplasmic Reticulum and Requires a Vma12p\/Vma22p Assembly Complex"],"prefix":"10.1083","volume":"142","author":[{"given":"Laurie A.","family":"Graham","sequence":"first","affiliation":[{"name":"Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kathryn J.","family":"Hill","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tom H.","family":"Stevens","sequence":"additional","affiliation":[{"name":"Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"291","published-online":{"date-parts":[[1998,7,13]]},"reference":[{"key":"2023072207473055800_B1","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1002\/yea.320090208","article-title":"The VPH2 gene encodes a 25 kD protein required for activity of the yeast vacuolar H+-ATPase","volume":"9","author":"Bachhawat","year":"1993","journal-title":"Yeast"},{"key":"2023072207473055800_B2","doi-asserted-by":"crossref","first-page":"12749","DOI":"10.1016\/S0021-9258(18)31452-2","article-title":"The Saccharomyces cerevisiae VMA6 gene encodes the 36-kD subunit of the vacuolar H+-ATPase membrane sector","volume":"268","author":"Bauerle","year":"1993","journal-title":"J Biol Chem"},{"key":"2023072207473055800_B3","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/S0962-8924(97)01020-9","article-title":"ER-associated and proteosome-mediated protein degradation: how two topologically restricted events came together","volume":"7","author":"Brodsky","year":"1997","journal-title":"Trends Cell Biol"},{"key":"2023072207473055800_B4","doi-asserted-by":"crossref","first-page":"5844","DOI":"10.1074\/jbc.271.10.5844","article-title":"A novel function for the second C2 domain of synaptotagamin; 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