Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1990 Dec;94(4):1874–1881. doi: 10.1104/pp.94.4.1874

Molecular Cloning of Tomato Plasma Membrane H+-ATPase 1

Nicholas N Ewing 1,2, Larry E Wimmers 1,2,2, David J Meyer 1,2, Roger T Chetelat 1,2, Alan B Bennett 1,2
PMCID: PMC1077467  PMID: 16667929

Abstract

Two cDNA clones (LHA1 and LHA2) from tomato (Lycopersicon esculentum) which likely encode isoforms of the plasma membrane H+-ATPase were isolated. The longest cDNA (3229 base pairs), LHA1, comprises an open reading frame that encodes a 956 amino acid, 105 kilodalton polypeptide with several potential transmembrane domains. In vitro transcription and translation of LHA1 yields a major translation product of approximately 100 kilodaltons that is immunoprecipitable with antiserum to the corn root plasma membrane H+-ATPase. LHA2 encodes a portion of a coding sequence that is 96% identical to LHA1, suggesting that LHA2 encodes an isoform of the H+-ATPase. Genomic DNA gel blot analysis indicates that both LHA1 and LHA2 hybridize to a common set of six to eight restriction fragments at moderate stringency and to single distinct fragments at high stringency. LHA1 and LHA2 map to distinct sites on chromosomes three and six, respectively. RNA gel blot analysis indicates that both LHA1 and LHA2 hybridize to 3.4 kilobase pair transcripts present in both leaves and roots, although the LHA2 transcript is relatively more abundant in leaves than in roots. These results indicate that in tomato as many as six to eight genes may encode the plasma membrane H+-ATPase, two of which are expressed at the level of mRNA in both roots and leaves.

Full text

PDF
1874

Images in this article

1878Figure 3
on p.1878
1878Figure 4
on p.1878
1879Figure 5
on p.1879
1880Figure 7
on p.1880

Selected References

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

  1. Alexander D. C. An efficient vector-primer cDNA cloning system. Methods Enzymol. 1987;154:41–64. doi: 10.1016/0076-6879(87)54069-1. [DOI] [PubMed] [Google Scholar]
  2. Anthon G. E., Spanswick R. M. Purification and properties of the h-translocating ATPase from the plasma membrane of tomato roots. Plant Physiol. 1986 Aug;81(4):1080–1085. doi: 10.1104/pp.81.4.1080. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boutry M., Michelet B., Goffeau A. Molecular cloning of a family of plant genes encoding a protein homologous to plasma membrane H+-translocating ATPases. Biochem Biophys Res Commun. 1989 Jul 31;162(2):567–574. doi: 10.1016/0006-291x(89)92348-6. [DOI] [PubMed] [Google Scholar]
  5. Briskin D. P., Leonard R. T. Partial characterization of a phosphorylated intermediate associated with the plasma membrane ATPase of corn roots. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6922–6926. doi: 10.1073/pnas.79.22.6922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Briskin D. P., Thornley W. R., Roti-Roti J. L. Target molecular size of the red beet plasma membrane ATPase. Plant Physiol. 1985 Jul;78(3):642–644. doi: 10.1104/pp.78.3.642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cathala G., Savouret J. F., Mendez B., West B. L., Karin M., Martial J. A., Baxter J. D. A method for isolation of intact, translationally active ribonucleic acid. DNA. 1983;2(4):329–335. doi: 10.1089/dna.1983.2.329. [DOI] [PubMed] [Google Scholar]
  8. Dellapenna D., Alexander D. C., Bennett A. B. Molecular cloning of tomato fruit polygalacturonase: Analysis of polygalacturonase mRNA levels during ripening. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6420–6424. doi: 10.1073/pnas.83.17.6420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hager K. M., Mandala S. M., Davenport J. W., Speicher D. W., Benz E. J., Jr, Slayman C. W. Amino acid sequence of the plasma membrane ATPase of Neurospora crassa: deduction from genomic and cDNA sequences. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7693–7697. doi: 10.1073/pnas.83.20.7693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Harper J. F., Surowy T. K., Sussman M. R. Molecular cloning and sequence of cDNA encoding the plasma membrane proton pump (H+-ATPase) of Arabidopsis thaliana. Proc Natl Acad Sci U S A. 1989 Feb;86(4):1234–1238. doi: 10.1073/pnas.86.4.1234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  12. MacLennan D. H., Brandl C. J., Korczak B., Green N. M. Amino-acid sequence of a Ca2+ + Mg2+-dependent ATPase from rabbit muscle sarcoplasmic reticulum, deduced from its complementary DNA sequence. Nature. 1985 Aug 22;316(6030):696–700. doi: 10.1038/316696a0. [DOI] [PubMed] [Google Scholar]
  13. Murray M. G., Thompson W. F. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 1980 Oct 10;8(19):4321–4325. doi: 10.1093/nar/8.19.4321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nevins J. R., Wilson M. C. Regulation of adenovirus-2 gene expression at the level of transcriptional termination and RNA processing. Nature. 1981 Mar 12;290(5802):113–118. doi: 10.1038/290113a0. [DOI] [PubMed] [Google Scholar]
  15. Oleski N. A., Bennett A. B. H-ATPase Activity from Storage Tissue of Beta vulgaris: IV. N,N'-Dicyclohexylcarbodiimide Binding and Inhibition of the Plasma Membrane H-ATPase. Plant Physiol. 1987 Mar;83(3):569–572. doi: 10.1104/pp.83.3.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pardo J. M., Serrano R. Structure of a plasma membrane H+-ATPase gene from the plant Arabidopsis thaliana. J Biol Chem. 1989 May 25;264(15):8557–8562. [PubMed] [Google Scholar]
  17. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Serrano R., Kielland-Brandt M. C., Fink G. R. Yeast plasma membrane ATPase is essential for growth and has homology with (Na+ + K+), K+- and Ca2+-ATPases. Nature. 1986 Feb 20;319(6055):689–693. doi: 10.1038/319689a0. [DOI] [PubMed] [Google Scholar]
  20. Shull G. E., Greeb J. Molecular cloning of two isoforms of the plasma membrane Ca2+-transporting ATPase from rat brain. Structural and functional domains exhibit similarity to Na+,K+- and other cation transport ATPases. J Biol Chem. 1988 Jun 25;263(18):8646–8657. [PubMed] [Google Scholar]
  21. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  22. Suiter K. A., Wendel J. F., Case J. S. LINKAGE-1: a PASCAL computer program for the detection and analysis of genetic linkage. J Hered. 1983 May-Jun;74(3):203–204. doi: 10.1093/oxfordjournals.jhered.a109766. [DOI] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES