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. 1995 Jun 20;92(13):6087–6091. doi: 10.1073/pnas.92.13.6087

Alternative splicing generates a second isoform of the catalytic A subunit of the vacuolar H(+)-ATPase.

N Hernando 1, M Bartkiewicz 1, P Collin-Osdoby 1, P Osdoby 1, R Baron 1
PMCID: PMC41647  PMID: 7597085

Abstract

We have identified a second isoform of the catalytic A subunit of the vacuolar H+ pump in chicken osteoclasts. In this isoform (A2) a 72-bp cassette replaces a 90-bp cassette present in the classical A1 isoform. The A1-specific cassette encodes a region of the protein that contains one of the three ATP-binding consensus sequences (the P-loop) identified in this polypeptide, as well as the pharmacologically relevant Cys254. In contrast, the A2-specific cassette does not contain any of these features. These two isoforms, which appear to be ubiquitously expressed, are encoded by a single gene and are generated by alternative splicing of two mutually exclusive exons. The alternative RNA processing involves the recognition of a single site, the boundary between the A2- and A1-specific exons, as either acceptor (in A1) or donor (in A2) splice site.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arai H., Berne M., Terres G., Terres H., Puopolo K., Forgac M. Subunit composition and ATP site labeling of the coated vesicle proton-translocating adenosinetriphosphatase. Biochemistry. 1987 Oct 20;26(21):6632–6638. doi: 10.1021/bi00395a011. [DOI] [PubMed] [Google Scholar]
  2. Baron R., Neff L., Louvard D., Courtoy P. J. Cell-mediated extracellular acidification and bone resorption: evidence for a low pH in resorbing lacunae and localization of a 100-kD lysosomal membrane protein at the osteoclast ruffled border. J Cell Biol. 1985 Dec;101(6):2210–2222. doi: 10.1083/jcb.101.6.2210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beachy P. A., Helfand S. L., Hogness D. S. Segmental distribution of bithorax complex proteins during Drosophila development. Nature. 1985 Feb 14;313(6003):545–551. doi: 10.1038/313545a0. [DOI] [PubMed] [Google Scholar]
  4. Billecocq A., Emanuel J. R., Levenson R., Baron R. 1 alpha,25-dihydroxyvitamin D3 regulates the expression of carbonic anhydrase II in nonerythroid avian bone marrow cells. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6470–6474. doi: 10.1073/pnas.87.16.6470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blair H. C., Teitelbaum S. L., Ghiselli R., Gluck S. Osteoclastic bone resorption by a polarized vacuolar proton pump. Science. 1989 Aug 25;245(4920):855–857. doi: 10.1126/science.2528207. [DOI] [PubMed] [Google Scholar]
  6. Boggs R. T., Gregor P., Idriss S., Belote J. M., McKeown M. Regulation of sexual differentiation in D. melanogaster via alternative splicing of RNA from the transformer gene. Cell. 1987 Aug 28;50(5):739–747. doi: 10.1016/0092-8674(87)90332-1. [DOI] [PubMed] [Google Scholar]
  7. Bowman E. J., Tenney K., Bowman B. J. Isolation of genes encoding the Neurospora vacuolar ATPase. Analysis of vma-1 encoding the 67-kDa subunit reveals homology to other ATPases. J Biol Chem. 1988 Oct 5;263(28):13994–14001. [PubMed] [Google Scholar]
  8. Brown D., Gluck S., Hartwig J. Structure of the novel membrane-coating material in proton-secreting epithelial cells and identification as an H+ATPase. J Cell Biol. 1987 Oct;105(4):1637–1648. doi: 10.1083/jcb.105.4.1637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chatterjee D., Chakraborty M., Leit M., Neff L., Jamsa-Kellokumpu S., Fuchs R., Baron R. Sensitivity to vanadate and isoforms of subunits A and B distinguish the osteoclast proton pump from other vacuolar H+ ATPases. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6257–6261. doi: 10.1073/pnas.89.14.6257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  11. David P., Baron R. The catalytic cycle of the vacuolar H(+)-ATPase. Comparison of proton transport in kidney- and osteoclast-derived vesicles. J Biol Chem. 1994 Dec 2;269(48):30158–30163. [PubMed] [Google Scholar]
  12. Dupont F. M., Tanaka C. K., Hurkman W. J. separation and Immunological Characterization of Membrane Fractions from Barley Roots. Plant Physiol. 1988 Mar;86(3):717–724. doi: 10.1104/pp.86.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Feng Y., Forgac M. Inhibition of vacuolar H(+)-ATPase by disulfide bond formation between cysteine 254 and cysteine 532 in subunit A. J Biol Chem. 1994 May 6;269(18):13224–13230. [PubMed] [Google Scholar]
  14. Forgac M. Structure and function of vacuolar class of ATP-driven proton pumps. Physiol Rev. 1989 Jul;69(3):765–796. doi: 10.1152/physrev.1989.69.3.765. [DOI] [PubMed] [Google Scholar]
  15. Gogarten J. P., Kibak H., Dittrich P., Taiz L., Bowman E. J., Bowman B. J., Manolson M. F., Poole R. J., Date T., Oshima T. Evolution of the vacuolar H+-ATPase: implications for the origin of eukaryotes. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6661–6665. doi: 10.1073/pnas.86.17.6661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Green M. R. Biochemical mechanisms of constitutive and regulated pre-mRNA splicing. Annu Rev Cell Biol. 1991;7:559–599. doi: 10.1146/annurev.cb.07.110191.003015. [DOI] [PubMed] [Google Scholar]
  17. Gräf R., Novak F. J., Harvey W. R., Wieczorek H. Cloning and sequencing of cDNA encoding the putative insect plasma membrane V-ATPase subunit A. FEBS Lett. 1992 Mar 30;300(2):119–122. doi: 10.1016/0014-5793(92)80177-i. [DOI] [PubMed] [Google Scholar]
  18. Hemken P., Guo X. L., Wang Z. Q., Zhang K., Gluck S. Immunologic evidence that vacuolar H+ ATPases with heterogeneous forms of Mr = 31,000 subunit have different membrane distributions in mammalian kidney. J Biol Chem. 1992 May 15;267(14):9948–9957. [PubMed] [Google Scholar]
  19. Hirata R., Ohsumk Y., Nakano A., Kawasaki H., Suzuki K., Anraku Y. Molecular structure of a gene, VMA1, encoding the catalytic subunit of H(+)-translocating adenosine triphosphatase from vacuolar membranes of Saccharomyces cerevisiae. J Biol Chem. 1990 Apr 25;265(12):6726–6733. [PubMed] [Google Scholar]
  20. Kane P. M. Biogenesis of the yeast vacuolar H(+)-ATPase. J Exp Biol. 1992 Nov;172:93–103. doi: 10.1242/jeb.172.1.93. [DOI] [PubMed] [Google Scholar]
  21. Mellman I., Fuchs R., Helenius A. Acidification of the endocytic and exocytic pathways. Annu Rev Biochem. 1986;55:663–700. doi: 10.1146/annurev.bi.55.070186.003311. [DOI] [PubMed] [Google Scholar]
  22. Nanda A., Gukovskaya A., Tseng J., Grinstein S. Activation of vacuolar-type proton pumps by protein kinase C. Role in neutrophil pH regulation. J Biol Chem. 1992 Nov 15;267(32):22740–22746. [PubMed] [Google Scholar]
  23. Narasimhan M. L., Binzel M. L., Perez-Prat E., Chen Z., Nelson D. E., Singh N. K., Bressan R. A., Hasegawa P. M. NaCl Regulation of Tonoplast ATPase 70-Kilodalton Subunit mRNA in Tobacco Cells. Plant Physiol. 1991 Oct;97(2):562–568. doi: 10.1104/pp.97.2.562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nelson H., Nelson N. Disruption of genes encoding subunits of yeast vacuolar H(+)-ATPase causes conditional lethality. Proc Natl Acad Sci U S A. 1990 May;87(9):3503–3507. doi: 10.1073/pnas.87.9.3503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nelson N., Taiz L. The evolution of H+-ATPases. Trends Biochem Sci. 1989 Mar;14(3):113–116. doi: 10.1016/0968-0004(89)90134-5. [DOI] [PubMed] [Google Scholar]
  26. Nelson R. D., Guo X. L., Masood K., Brown D., Kalkbrenner M., Gluck S. Selectively amplified expression of an isoform of the vacuolar H(+)-ATPase 56-kilodalton subunit in renal intercalated cells. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3541–3545. doi: 10.1073/pnas.89.8.3541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pan Y. X., Xu J., Strasser J. E., Howell M., Dean G. E. Structure and expression of subunit A from the bovine chromaffin cell vacuolar ATPase. FEBS Lett. 1991 Nov 18;293(1-2):89–92. doi: 10.1016/0014-5793(91)81158-5. [DOI] [PubMed] [Google Scholar]
  28. Peng S. B., Crider B. P., Xie X. S., Stone D. K. Alternative mRNA splicing generates tissue-specific isoforms of 116-kDa polypeptide of vacuolar proton pump. J Biol Chem. 1994 Jun 24;269(25):17262–17266. [PubMed] [Google Scholar]
  29. Puopolo K., Kumamoto C., Adachi I., Forgac M. A single gene encodes the catalytic "A" subunit of the bovine vacuolar H(+)-ATPase. J Biol Chem. 1991 Dec 25;266(36):24564–24572. [PubMed] [Google Scholar]
  30. Puopolo K., Kumamoto C., Adachi I., Magner R., Forgac M. Differential expression of the "B" subunit of the vacuolar H(+)-ATPase in bovine tissues. J Biol Chem. 1992 Feb 25;267(6):3696–3706. [PubMed] [Google Scholar]
  31. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Saraste M., Sibbald P. R., Wittinghofer A. The P-loop--a common motif in ATP- and GTP-binding proteins. Trends Biochem Sci. 1990 Nov;15(11):430–434. doi: 10.1016/0968-0004(90)90281-f. [DOI] [PubMed] [Google Scholar]
  33. Smith C. W., Patton J. G., Nadal-Ginard B. Alternative splicing in the control of gene expression. Annu Rev Genet. 1989;23:527–577. doi: 10.1146/annurev.ge.23.120189.002523. [DOI] [PubMed] [Google Scholar]
  34. Takeyasu K., Lemas V., Fambrough D. M. Stability of Na(+)-K(+)-ATPase alpha-subunit isoforms in evolution. Am J Physiol. 1990 Oct;259(4 Pt 1):C619–C630. doi: 10.1152/ajpcell.1990.259.4.C619. [DOI] [PubMed] [Google Scholar]
  35. Umemoto N., Ohya Y., Anraku Y. VMA11, a novel gene that encodes a putative proteolipid, is indispensable for expression of yeast vacuolar membrane H(+)-ATPase activity. J Biol Chem. 1991 Dec 25;266(36):24526–24532. [PubMed] [Google Scholar]
  36. Vänänen H. K., Karhukorpi E. K., Sundquist K., Wallmark B., Roininen I., Hentunen T., Tuukkanen J., Lakkakorpi P. Evidence for the presence of a proton pump of the vacuolar H(+)-ATPase type in the ruffled borders of osteoclasts. J Cell Biol. 1990 Sep;111(3):1305–1311. doi: 10.1083/jcb.111.3.1305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Walker J. E., Saraste M., Runswick M. J., Gay N. J. Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J. 1982;1(8):945–951. doi: 10.1002/j.1460-2075.1982.tb01276.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ziff E. B. Transcription and RNA processing by the DNA tumour viruses. Nature. 1980 Oct 9;287(5782):491–499. doi: 10.1038/287491a0. [DOI] [PubMed] [Google Scholar]
  39. Zimniak L., Dittrich P., Gogarten J. P., Kibak H., Taiz L. The cDNA sequence of the 69-kDa subunit of the carrot vacuolar H+-ATPase. Homology to the beta-chain of F0F1-ATPases. J Biol Chem. 1988 Jul 5;263(19):9102–9112. [PubMed] [Google Scholar]

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