Disassembly and Reassembly of the Yeast Vacuolar H+-ATPase in Vivo

Abstract

The vacuolar H+-ATPase of the yeast Saccharomyces cerevisiae is composed of a complex of peripheral subunits (the V1 sector) attached to an integral membrane complex (the V0 sector). In the experiments described here, attachment of the V1 to the V0 sector was assessed in wild-type cells under a variety of growth conditions. Depriving the yeast cells of glucose, even for as little as 5 min, caused dissociation of approximately 70% of the assembled enzyme complexes into separate V1 and V0 subcomplexes. Restoration of glucose induced rapid and efficient reassembly of the enzyme from the previously synthesized subcomplexes. Indirect immunofluorescence microscopy and subcellular fractionation revealed detachment of the peripheral subunits from the vacuolar membrane in the absence of glucose, followed by reattachment in the presence of glucose. Rapid dissociation of vacuolar H+-ATPases could also be triggered by shifting cells into a variety of other carbon sources, and reassembly could be generated by addition of glucose. Disassembly and reassembly of vacuolar H+-ATPases in vivo may be a means of regulating organelle acidification in response to extracellular conditions, or a mechanism for assembling alternate complexes of vacuolar H+-ATPases in different intracellular compartments. The vacuolar H+-ATPase of the yeast Saccharomyces cerevisiae is composed of a complex of peripheral subunits (the V1 sector) attached to an integral membrane complex (the V0 sector). In the experiments described here, attachment of the V1 to the V0 sector was assessed in wild-type cells under a variety of growth conditions. Depriving the yeast cells of glucose, even for as little as 5 min, caused dissociation of approximately 70% of the assembled enzyme complexes into separate V1 and V0 subcomplexes. Restoration of glucose induced rapid and efficient reassembly of the enzyme from the previously synthesized subcomplexes. Indirect immunofluorescence microscopy and subcellular fractionation revealed detachment of the peripheral subunits from the vacuolar membrane in the absence of glucose, followed by reattachment in the presence of glucose. Rapid dissociation of vacuolar H+-ATPases could also be triggered by shifting cells into a variety of other carbon sources, and reassembly could be generated by addition of glucose. Disassembly and reassembly of vacuolar H+-ATPases in vivo may be a means of regulating organelle acidification in response to extracellular conditions, or a mechanism for assembling alternate complexes of vacuolar H+-ATPases in different intracellular compartments. Vacuolar proton-translocating ATPases (H+-ATPases)1 1The abbreviations used are: H+-ATPaseproton-translocating ATPaseYEPDyeast extract-peptone-2% dextrose mediumYEP1% yeast extract-2% peptone medium without dextroseSD-Metsupplemented minimal medium lacking methionineDSPdithiobis(succinimidylpropionate)PBSphosphate-buffered saline. 1The abbreviations used are: H+-ATPaseproton-translocating ATPaseYEPDyeast extract-peptone-2% dextrose mediumYEP1% yeast extract-2% peptone medium without dextroseSD-Metsupplemented minimal medium lacking methionineDSPdithiobis(succinimidylpropionate)PBSphosphate-buffered saline. are multisubunit complexes, found in all eukaryotic cells, that acidify one or more intracellular compartments(1Forgac M. Physiol. Rev. 1989; 69: 765-796Crossref PubMed Scopus (479) Google Scholar). Vacuolar H+-ATPases from plants, animals, and fungi are very similar. All consist of a V1 sector, which is a complex of peripheral membrane proteins containing the ATP-binding sites, and a V0 sector, comprising a complex of integral membrane proteins containing the proton pore(2Sze H. Ward J.M. Lai S. J. Bioenerg. Biomemb. 1992; 24: 371-381Crossref PubMed Scopus (180) Google Scholar, 3Forgac M. J. Bioenerg. Biomemb. 1992; 24: 341-350Crossref PubMed Scopus (50) Google Scholar, 4Kane P.M. Stevens T.H. J. Bioenerg. Biomemb. 1992; 24: 383-393Crossref PubMed Scopus (56) Google Scholar). The vacuolar H+-ATPases share sequence and structural similarities with the F1F0-ATPases of mitochondria, chloroplasts, and bacteria, and these similarities are believed to reflect an evolutionary relationship(5Nelson N. Taiz L. Trends Biochem. Sci. 1989; 14: 113-116Abstract Full Text PDF PubMed Scopus (244) Google Scholar). However, unlike the F1F0-ATPases, which can be separated into a soluble F1 complex that is capable of ATP hydrolysis(6Penefsky H.S. Cross R.L. Adv. Enzymol. Relat. Areas Mol. Biol. 1990; 64: 173-214Google Scholar), and a membrane-bound F0 complex that appears to function as a proton channel(7Lill H. Engelbrecht S. Schonknecht G. Junge W. Eur. J. Biochem. 1986; 160: 627-634Crossref PubMed Scopus (34) Google Scholar), a variety of biochemical studies indicate that the V1 sector of the vacuolar H+-ATPases does not retain its ATPase activity when detached from the membrane(8Bowman B.J. Dschida W.J. Harris T. Bowman E.J. J. Biol. Chem. 1989; 264: 15606-15612Abstract Full Text PDF PubMed Google Scholar, 9Kane P.M. Yamashiro C.T. Stevens T.H. J. Biol. Chem. 1989; 264: 19236-19244Abstract Full Text PDF PubMed Google Scholar, 10Moriyama Y. Nelson N. J. Biol. Chem. 1989; 264: 3577-3582Abstract Full Text PDF PubMed Google Scholar, 11Puopolo K. Forgac M. J. Biol. Chem. 1990; 265: 14836-14841Abstract Full Text PDF PubMed Google Scholar, 12Ward J.M. Sze H. Plant Physiol. 1991; 99: 170-179Crossref Scopus (68) Google Scholar), and the V0 complex is not an open proton pore(12Ward J.M. Sze H. Plant Physiol. 1991; 99: 170-179Crossref Scopus (68) Google Scholar, 13Zhang J. Myers M. Forgac M. J. Biol. Chem. 1992; 267: 9773-9778Abstract Full Text PDF PubMed Google Scholar). proton-translocating ATPase yeast extract-peptone-2% dextrose medium 1% yeast extract-2% peptone medium without dextrose supplemented minimal medium lacking methionine dithiobis(succinimidylpropionate) phosphate-buffered saline. proton-translocating ATPase yeast extract-peptone-2% dextrose medium 1% yeast extract-2% peptone medium without dextrose supplemented minimal medium lacking methionine dithiobis(succinimidylpropionate) phosphate-buffered saline. Studies of the assembly of the yeast vacuolar H+-ATPase have shown that mutants lacking one subunit of the enzyme complex can still assemble the V1 complex in the cytoplasm under conditions where the V0 complex is not assembled, and the V0 complex can be assembled and transported to the vacuole in the absence of an assembled V1 complex(14Doherty R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar, 15Kane P.M. Kuehn M.C. Howald-Stevenson I. Stevens T.H. J. Biol. Chem. 1992; 267: 447-454Abstract Full Text PDF PubMed Google Scholar, 16Noumi T. Beltran C. Nelson H. Nelson N. Proc. Natl. Acad. Sci. U. S. A. 1991; 88: 1938-1942Crossref PubMed Scopus (128) Google Scholar, 17Umemoto N. Yoshihisa T. Hirata R. Anraku Y. J. Biol. Chem. 1990; 265: 18447-18453Abstract Full Text PDF PubMed Google Scholar). Independent V1 and V0 complexes were also seen in wild-type yeast cells, but it was not clear whether these complexes were assembly intermediates, products of breakdown of the vacuolar or of the biochemical R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar). the dissociation of the wild-type V1 complex from the V0 complex in the and reassembly of the complex and in response to in growth conditions, that these could a in of vacuolar H+-ATPase activity or in of the was from was from was from were from All used for immunofluorescence microscopy and were from All other were from The wild-type yeast was used in all were as described in Scholar, C.T. Kane P.M. Stevens T.H. Mol. Biol. 1990; Scopus Google Scholar). were under conditions as R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar), with the were to in supplemented minimal medium lacking methionine that glucose and by for with of were in of and for the with of the of the methionine and were to a of and the cells were by with a the were in the medium to the as for and for the and were were in phosphate-buffered containing 1% and a 5 was as in to a of min, an of containing a of the was to the to the and min, were with as were as R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar), that 5 of P.M. Yamashiro C.T. Stevens T.H. J. Biol. Chem. 1989; 264: 19236-19244Abstract Full Text PDF PubMed Google and of containing 5 were of a containing proteins were and by and as R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar). In were by the to the and a In the shown in yeast cells were to and with as described that was used as the carbon of glucose. methionine and were to the but a of and glucose a of were to the medium the and in assembly of complexes in cells in in a In the absence of the a of with the of the could be from wild-type cells by the The subunit to be without even it could be with the complex by other R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar). of of the subunit with the wild-type complex and more different In the presence of proteins in addition to the and subunits of the ATPase are by the a and a that were previously in the absence of R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar), as as a and of these was of the but the of in not to under conditions for and reassembly as the and proteins were to in yeast dextrose to approximately in or yeast peptone medium without and the of cells were for in the and Yamashiro C.T. Stevens T.H. Enzymol. 1991; PubMed Scopus Google Scholar). and were used a and and the was as Yamashiro C.T. Stevens T.H. Enzymol. 1991; PubMed Scopus Google Scholar). to to in containing or containing were to and as described that all were in the presence of the carbon from approximately cells were in of the medium containing and for In the was for in the presence of glucose to the cells were the in of containing the used for was to a of and the was for The was by addition of of followed by for addition of of containing the the was in a with of the The was for in an and were by as R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar). Vacuolar were as Yamashiro C.T. Stevens T.H. Enzymol. 1991; PubMed Scopus Google Scholar), that were with in containing or containing the the were by ATPase of were the enzyme of C. PubMed Scopus Google Scholar), and a J. Biol. Chem. Full Text PDF PubMed Google was used to In to the of ATPase activity for vacuolar were with for and the the ATPase of with and without with was as the vacuolar ATPase is a of S. Bowman E.J. A. A. K. 1993; PubMed Scopus Google whether the of assembly of the yeast vacuolar H+-ATPase is in response to in extracellular conditions, yeast were with for and to different containing methionine for a to that for min, all of the previously subunits of the yeast vacuolar H+-ATPase can be with the subunit of the enzyme and is little in the of subunits a in the medium that assembly of the yeast vacuolar H+-ATPase is the and that the assembled complexes are the complexes were from cells in medium to 5 to or containing conditions were yeast mutants lacking vacuolar H+-ATPase activity to in medium to or containing but are to medium to that for the vacuolar H+-ATPase under these different growth conditions However, these indicate that the different not assembled vacuolar H+-ATPase In assembly of the yeast vacuolar H+-ATPase was by the cells to medium lacking glucose for from cells in medium without glucose of the and appears to be in the of the subunit in was not seen as The of the subunit in the experiments shown in and was more The and proteins that with the subunit in medium lacking glucose have as of the peripheral V1 sector of the vacuolar with the subunit in The and subunits have shown to be integral membrane P.M. Kuehn M.C. Howald-Stevenson I. Stevens T.H. J. Biol. Chem. 1992; 267: 447-454Abstract Full Text PDF PubMed Google Scholar, 16Noumi T. Beltran C. Nelson H. Nelson N. Proc. Natl. Acad. Sci. U. S. A. 1991; 88: 1938-1942Crossref PubMed Scopus (128) Google Scholar, 17Umemoto N. Yoshihisa T. Hirata R. Anraku Y. J. Biol. Chem. 1990; 265: 18447-18453Abstract Full Text PDF PubMed Google Scholar), and the subunit appears to be with these subunits even it is not an integral membrane subunit and reassembly of the vacuolar H+-ATPase not cells were to and with for as described under were or for in to 5 in to 5 and in to 5 and or in to followed by an with glucose 5 and In the medium All of the and were and proteins were by and H+-ATPase subunits are as the complexes from the by the which an the subunit that appears to be when the V1 subunits are not P.M. Kuehn M.C. Howald-Stevenson I. Stevens T.H. J. Biol. Chem. 1992; 267: 447-454Abstract Full Text PDF PubMed Google Scholar). used to P.M. Kuehn M.C. Howald-Stevenson I. Stevens T.H. J. Biol. Chem. 1992; 267: 447-454Abstract Full Text PDF PubMed Google and R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google V0 complexes under conditions where the peripheral V1 complex to the The a complex containing the and in addition to other proteins that have not from wild-type cells that the cells a of assembled V0 complexes that are not with the V1 subunits the in glucose medium is an in the of and subunits by the The in indicate in medium lacking glucose caused dissociation of the peripheral V1 sector of the yeast vacuolar H+-ATPase from the integral membrane V0 and the subunits of sector, with the of the The dissociation of the peripheral V1 sector be to the ATPase activity of vacuolar vacuolar membrane were from in the presence or absence of glucose from in medium containing glucose of vacuolar ATPase activity of ATP and vacuolar ATPase activity is as the ATPase activity by The in the absence of glucose containing vacuolar ATPase that of in the absence of glucose in a of of the vacuolar ATPase activity from the The presence of assembled V1 and V0 in the cells in medium lacking glucose that the complexes for whether of the vacuolar H+-ATPase is a with medium lacking glucose for and glucose for and the shown in and the assembled vacuolar H+-ATPase complexes were in medium without glucose. However, glucose was to the cells for and the complexes were from from cells with glucose the and and reassembly of the Disassembly in medium lacking glucose, and reassembly when glucose was to All of the subunits are in complexes the of subunits is not for it is still that other proteins in or reassembly be in the medium and and the the of was shown previously to yeast S. R. Full Text PDF PubMed Scopus Google of and in that does not to have a the assembled vacuolar H+-ATPase complexes, and and that and reassembly can in the absence of In to a of the of and reassembly of the vacuolar H+-ATPase in the presence and absence of glucose, the of the and subunits the complexes by the and the complexes by the subunits was a The are shown In cells for in the presence of glucose, of the of these V0 subunits by was with the V1 that of the V0 subunits are found as of assembled ATPase However, when the cells were to a in the absence of glucose, of the subunits were by the and and were by the that the subunits are found in V0 complexes not containing V1 of glucose the seen cells were of glucose. Disassembly and reassembly of the yeast vacuolar H+-ATPase were also by immunofluorescence and the are shown in as when cells are under and Indirect immunofluorescence microscopy with the which the peripheral subunit of the of the V1 sector with the vacuolar membrane in yeast cells in the presence of glucose the cells have of glucose for min, the of the in addition to of the vacuolar membrane Restoration of glucose to the cells in of and of vacuolar membrane are with the from described in of and reassembly of the vacuolar H+-ATPase complex was from immunofluorescence microscopy the and was previously shown to little or of wild-type cells in the presence of P.M. Kuehn M.C. Howald-Stevenson I. Stevens T.H. J. Biol. Chem. 1992; 267: 447-454Abstract Full Text PDF PubMed Google Scholar), and in appears to be very little of cells with glucose or cells of glucose that the glucose However, the the vacuolar membrane in cells of glucose that the the subunit is in these experiments mutants that to assemble the peripheral subunits the vacuolar membrane P.M. Kuehn M.C. Howald-Stevenson I. Stevens T.H. J. Biol. Chem. 1992; 267: 447-454Abstract Full Text PDF PubMed Google Scholar), that of the of the peripheral membrane subunits from the integral membrane The microscopy in and also that and reassembly in yeast cells were in the presence and absence of glucose and were to in these immunofluorescence microscopy of the subunit in yeast cells under conditions. were as but were with to the V0 sector when it is not with the V1 were under and or and The dissociation of the yeast vacuolar H+-ATPase in medium lacking carbon that also be rapid in the ATPase when cells were different carbon cells and for in glucose were to different carbon for a and the are shown in of the complexes in and in of the vacuolar all of these conditions, the and subunits to be from the complexes by the and of the and subunits in the V0 by the of glucose a of for could reassembly of the vacuolar H+-ATPase from previously synthesized V1 and V0 in all of these carbon and The of of the vacuolar H+-ATPase were by the in medium lacking glucose. the without the complexes were to seen in medium without glucose not the of reassembly were by of complexes of glucose to that of glucose for also to be a in glucose not of the may have as as to the or addition of glucose and from glucose and reassembly with glucose are in 5 In the and of under different conditions are were a and the of the and subunits by the is shown as a of assembly and The in glucose, and these that a of assembly of the subunit with the subunit appears to the However, of the in the absence of carbon or in the presence of or induced a rapid dissociation of the subunit from the complexes by the The and subunits were a under these conditions not The may have in in the other carbon sources, but for all of the was by the in the the in medium lacking glucose was not seen in other is also that the of is under of the different conditions. with the a in that carbon and as as and as subunit as a in glucose. The microscopy shown in indicate that and reassembly in the absence of glucose are not to the vacuolar H+-ATPase complexes synthesized the of the was by of the of the and subunits and from that under conditions, and with as described under The peripheral subunits were found in the from in glucose and even the to a of glucose to glucose However, the were in medium lacking glucose the peripheral subunits were found in the All of the experiments described in assembly of vacuolar H+-ATPase complexes that to assemble in However, the of the ATPase in a variety of different carbon that assembly of the enzyme growth these carbon could also be yeast cells were in containing or as carbon and the of the and subunits membrane and was by In cells in the and peripheral subunits to be in the that are not assembled or not the membrane In cells in approximately of the peripheral subunits are found in the that assembly or of the subunits the membrane is efficient in cells but more efficient in The presence of the peripheral subunits in the from cells in could from a in the of V0 sector to the V1 subunits that are for the V1 However, when from cells in are for in glucose, the and subunits are to the that are V0 subunits or complexes in the cells that to V1 subunits a in glucose. was by vacuolar H+-ATPase complexes growth and the assembly of the subunit into complexes a in or glucose. from cells in were for with in the presence of an of methionine was with or glucose. the of the and subunits by the as of complexes, as by is very little in the of the V0 subunits by the subunit a in that of these subunits can assemble into complexes in the are in V0 In addition of glucose for a of these subunits from V0 complexes to assembled the assembly of the V0 subunits into complexes is still the assembly shown in in glucose, it is clear that the V0 complexes in cells be to be into assembled complexes when the is The have clear for the of the yeast vacuolar H+-ATPase and also for enzyme or of vacuolar H+-ATPases in response to extracellular conditions. a for ATPases in which the V1 and V0 may be but are B.J. Dschida W.J. Harris T. Bowman E.J. J. Biol. Chem. 1989; 264: 15606-15612Abstract Full Text PDF PubMed Google Scholar, 9Kane P.M. Yamashiro C.T. Stevens T.H. J. Biol. Chem. 1989; 264: 19236-19244Abstract Full Text PDF PubMed Google Scholar, 10Moriyama Y. Nelson N. J. Biol. Chem. 1989; 264: 3577-3582Abstract Full Text PDF PubMed Google Scholar, 11Puopolo K. Forgac M. J. Biol. Chem. 1990; 265: 14836-14841Abstract Full Text PDF PubMed Google Scholar, 12Ward J.M. Sze H. Plant Physiol. 1991; 99: 170-179Crossref Scopus (68) Google Scholar, 13Zhang J. Myers M. Forgac M. J. Biol. Chem. 1992; 267: 9773-9778Abstract Full Text PDF PubMed Google Scholar, R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar). Independent V1 and V0 complexes can in mutants lacking one R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar), and a of have the presence of V1 and V0 in wild-type yeast cells as as in other J. Myers M. Forgac M. J. Biol. Chem. 1992; 267: 9773-9778Abstract Full Text PDF PubMed Google Scholar, R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar, 17Umemoto N. Yoshihisa T. Hirata R. Anraku Y. J. Biol. Chem. 1990; 265: 18447-18453Abstract Full Text PDF PubMed Google Scholar, M. Forgac M. J. Physiol. 1993; PubMed Scopus Google Scholar). for these is that assembled complexes are biochemical it is to the that dissociation for all of the V1 and V0 in subunit with different are seen by a of different under conditions, subcellular and immunofluorescence and in these have by in ATPase activity of In to dissociation of the complex a was also in the and fractionation In the presence of the the subunit of the complexes and the of the V1 subunits soluble and membrane previously in the absence of R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google Scholar), with the of the which could be in the presence of However, the of the V0 subunits with the V1 subunits and the of subunits in experiments that shown in were found to be more different experiments when the was In the absence of were experiments when very little of the V0 subunits were by the and experiments where all of the peripheral subunits were found in the from of whether the cells in glucose. of may have caused by of the subunit in the absence of experiments the ATPase have that complexes can in the absence of the these complexes are K. M. R. Forgac M. J. Biol. Chem. 1992; 267: Full Text PDF PubMed Google Scholar). the of the V1 and V0 subunits with a of and the for dissociation of complexes the for that the V1 and V0 of the ATPases can in The structural from these is that the V1 and V0 in be in a with assembled conditions that ATPase the and V1 subunits are as a complex to the the and V0 subunits assembled in the The subunit appears to be from in with R.D. Kane P.M. J. Biol. Chem. 1993; 268: 16845-16851Abstract Full Text PDF PubMed Google and biochemical yeast cells can vacuolar H+-ATPase complexes previously synthesized subunits in response to in extracellular conditions. in the absence of glucose, the integral V0 sector, the peripheral V1 sector, and the subunit all for the rapid of and reassembly and the of in the presence of indicate that activity for dissociation and of the complex be the assembled vacuolar H+-ATPase or in other proteins synthesized the The V1 and V0 described in yeast cells R.D. Kane P.M. J. Biol. 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The and of the Scholar). and glucose are used in but glucose and glucose is growth is a carbon and a still different of proteins for the variety of of shifting to these different carbon sources, the of the vacuolar H+-ATPase seen a to of these carbon is very to that seen in the absence of carbon of glucose to of these carbon a rapid of the V1 and V0 as of and also when cells are from or growth to rapid J.M. S. Trends Biochem. Sci. Full Text PDF PubMed Scopus Google Scholar, M. M. The and of the Scholar). The in assembly of the vacuolar H+-ATPase when cells are from glucose are rapid and can in the presence of a However, cells in or also of ATPase and rapid of carbon ATPase The and reassembly could a mechanism for regulating acidification of the yeast vacuole or other membrane attachment is for ATPase activity in as it is in the with the other peripheral from the vacuolar could proton and ATP it shown that rapid in seen of were caused by a of the V1 subunits from the membrane which in of ATPase activity and proton U. H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). is that the structural that and reassembly in the yeast also the dissociation of the vacuolar and it is to whether the intracellular could in is also that the yeast membrane H+-ATPase is by addition of glucose to cells in or growth and by intracellular acidification of growth in a as J.M. S. Trends Biochem. Sci. Full Text PDF PubMed Scopus Google Scholar, R. The and of the Scholar), and that is by in of the A. C. J. Biol. 1991; PubMed Scopus Google Scholar). of the membrane H+-ATPase when growth in glucose is as a mechanism of regulating R. The and of the Scholar), and an of assembled vacuolar H+-ATPases induced by addition of glucose could also in In cells V0 complexes, or of the V0 as in yeast and in other have J. Myers M. Forgac M. J. Biol. Chem. 1992; 267: 9773-9778Abstract Full Text PDF PubMed Google Scholar, J. Biol. Chem. Full Text PDF PubMed Google Scholar, A. T. J. J. Biol. Chem. 1992; 267: Full Text PDF PubMed Google Scholar, S. Sze H. J. Biol. Chem. 1991; Full Text PDF PubMed Google Scholar), the reassembly could the V1 complexes to different or the of the subunits of the of and reassembly into the structural the vacuolar H+-ATPase as as the for of organelle were with for and for an in the presence or absence of glucose, as and assembled complexes of the vacuolar H+-ATPase were the and and by as described under The of in of the and subunit by the was a The of subunit as of assembled complexes was by the by the to the by The by the the of the subunits in V0 complexes not assembled with V1 were with in medium containing for and for a of 5 or addition or glucose. and assembled complexes of the vacuolar H+-ATPase were the and and by as described under The of subunit by the as of assembled complexes was as described in of and subunits as of V0 complexes different in a for the of for with and for a of the