Skip to main content
The Plant Cell logoLink to The Plant Cell
. 1994 Oct;6(10):1375–1389. doi: 10.1105/tpc.6.10.1375

A plant plasma membrane proton-ATPase gene is regulated by development and environment and shows signs of a translational regulation.

B Michelet 1, M Lukaszewicz 1, V Dupriez 1, M Boutry 1
PMCID: PMC160527  PMID: 7994172

Abstract

A proton-pumping ATPase is present in the plasma membrane of plant cells where it sustains transport-related functions. This enzyme is encoded by a family of genes that shows signs of both transcriptional and post-transcriptional regulation. The regulation of pma1, one of the Nicotiana plumbaginifolia H+-ATPase genes, was characterized with the help of the beta-glucuronidase (gusA) receptor gene in transgenic plants. pma1 is active in the root epidermis, the stem cortex, and guard cells. This activity depends on developmental and growth conditions. For instance, pma1 activity in guard cells was strongly enhanced when the plant material (young seedlings or mature leaves) was incubated in liquid growth medium. pma1 is also expressed in several tissues of the reproductive organs where active transport is thought to occur but where scarcely any ATPase activity has been identified, namely in the tapetum, the pollen, the transmitting tissue, and the ovules. Several pma genes have a long 5'untranslated region (leader sequence) containing an upstream open reading frame (URF). Analysis of translational and transcriptional fusions with gusA in transgenic plants suggests that the pma1 leader sequence might activate translation of the main open reading frame, even though the URF is translated by a large majority of the scanning ribosomes. As confirmation, transient expression experiments showed that the pma1 leader causes a fourfold post-transcriptional increase of main open reading frame expression. Deletion of the URF by site-directed mutagenesis stimulated the main open reading frame translation 2.7-fold in an in vitro translational assay. These results are consistent with a regulatory mechanism involving translation reinitiation. Altogether, they suggest a fine, multilevel regulation of H+-ATPase activity in the plant.

Full Text

The Full Text of this article is available as a PDF (2.6 MB).

Selected References

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

  1. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boutry M., Chua N. H. A nuclear gene encoding the beta subunit of the mitochondrial ATP synthase in Nicotiana plumbaginifolia. EMBO J. 1985 Sep;4(9):2159–2165. doi: 10.1002/j.1460-2075.1985.tb03910.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Chaumont F., O'Riordan V., Boutry M. Protein transport into mitochondria is conserved between plant and yeast species. J Biol Chem. 1990 Oct 5;265(28):16856–16862. [PubMed] [Google Scholar]
  6. DeWitt N. D., Harper J. F., Sussman M. R. Evidence for a plasma membrane proton pump in phloem cells of higher plants. Plant J. 1991 Jul;1(1):121–128. doi: 10.1111/j.1365-313x.1991.00121.x. [DOI] [PubMed] [Google Scholar]
  7. Dente L., Cesareni G., Cortese R. pEMBL: a new family of single stranded plasmids. Nucleic Acids Res. 1983 Mar 25;11(6):1645–1655. doi: 10.1093/nar/11.6.1645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ewing N. N., Wimmers L. E., Meyer D. J., Chetelat R. T., Bennett A. B. Molecular Cloning of Tomato Plasma Membrane H-ATPase. Plant Physiol. 1990 Dec;94(4):1874–1881. doi: 10.1104/pp.94.4.1874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fütterer J., Hohn T. Role of an upstream open reading frame in the translation of polycistronic mRNAs in plant cells. Nucleic Acids Res. 1992 Aug 11;20(15):3851–3857. doi: 10.1093/nar/20.15.3851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gallie D. R., Walbot V. Identification of the motifs within the tobacco mosaic virus 5'-leader responsible for enhancing translation. Nucleic Acids Res. 1992 Sep 11;20(17):4631–4638. doi: 10.1093/nar/20.17.4631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Green P. J. Control of mRNA Stability in Higher Plants. Plant Physiol. 1993 Aug;102(4):1065–1070. doi: 10.1104/pp.102.4.1065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Harper J. F., Manney L., DeWitt N. D., Yoo M. H., Sussman M. R. The Arabidopsis thaliana plasma membrane H(+)-ATPase multigene family. Genomic sequence and expression of a third isoform. J Biol Chem. 1990 Aug 15;265(23):13601–13608. [PubMed] [Google Scholar]
  13. 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]
  14. Hinnebusch A. G. Novel mechanisms of translational control in Saccharomyces cerevisiae. Trends Genet. 1988 Jun;4(6):169–174. doi: 10.1016/0168-9525(88)90023-6. [DOI] [PubMed] [Google Scholar]
  15. Houlné G., Boutry M. Identification of an Arabidopsis thaliana gene encoding a plasma membrane H(+)-ATPase whose expression is restricted to anther tissue. Plant J. 1994 Mar;5(3):311–317. doi: 10.1111/j.1365-313x.1994.00311.x. [DOI] [PubMed] [Google Scholar]
  16. Jackson R. J. Cytoplasmic regulation of mRNA function: the importance of the 3' untranslated region. Cell. 1993 Jul 16;74(1):9–14. doi: 10.1016/0092-8674(93)90290-7. [DOI] [PubMed] [Google Scholar]
  17. Jefferson R. A., Kavanagh T. A., Bevan M. W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 1987 Dec 20;6(13):3901–3907. doi: 10.1002/j.1460-2075.1987.tb02730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kozak M. An analysis of vertebrate mRNA sequences: intimations of translational control. J Cell Biol. 1991 Nov;115(4):887–903. doi: 10.1083/jcb.115.4.887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Lohmer S., Maddaloni M., Motto M., Salamini F., Thompson R. D. Translation of the mRNA of the maize transcriptional activator Opaque-2 is inhibited by upstream open reading frames present in the leader sequence. Plant Cell. 1993 Jan;5(1):65–73. doi: 10.1105/tpc.5.1.65. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Moriau L., Bogaerts P., Jonniaux J. L., Boutry M. Identification and characterization of a second plasma membrane H(+)-ATPase gene subfamily in Nicotiana plumbaginifolia. Plant Mol Biol. 1993 Mar;21(6):955–963. doi: 10.1007/BF00023594. [DOI] [PubMed] [Google Scholar]
  23. Perez C., Michelet B., Ferrant V., Bogaerts P., Boutry M. Differential expression within a three-gene subfamily encoding a plasma membrane H(+)-ATPase in Nicotiana plumbaginifolia. J Biol Chem. 1992 Jan 15;267(2):1204–1211. [PubMed] [Google Scholar]
  24. Samuels A. L., Fernando M., Glass A. D. Immunofluorescent Localization of Plasma Membrane H-ATPase in Barley Roots and Effects of K Nutrition. Plant Physiol. 1992 Aug;99(4):1509–1514. doi: 10.1104/pp.99.4.1509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sussman M. R., Harper J. F. Molecular biology of the plasma membrane of higher plants. Plant Cell. 1989 Oct;1(10):953–960. doi: 10.1105/tpc.1.10.953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Uknes S., Dincher S., Friedrich L., Negrotto D., Williams S., Thompson-Taylor H., Potter S., Ward E., Ryals J. Regulation of pathogenesis-related protein-1a gene expression in tobacco. Plant Cell. 1993 Feb;5(2):159–169. doi: 10.1105/tpc.5.2.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Verwoerd T. C., Dekker B. M., Hoekema A. A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res. 1989 Mar 25;17(6):2362–2362. doi: 10.1093/nar/17.6.2362. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES