Abstract
The plasma membrane H+-ATPases in Arabidopsis thaliana represent the largest family of cation translocating P-type ATPases identified in plants or animals. We report here seven new isoforms, which were identified by polymerase chain reaction (PCR) amplification of genomic DNA. Amplifications were performed with degenerate primers corresponding to two short conserved sequence motifs (“CSDK” and “GDGV”) found in most P-type ATPases. A comparison was made of three CSDK-side primers, which were used either as totally degenerate mixtures or rendered less degenerate by substitution with deoxyinosine or fluorodeoxyuridine. Amplified genomic fragments were cloned, partially sequenced and shown to correspond to Arabidopsis genes by Southern blot analysis with gene-specific probes. One newly identified isoform, AHA10, was isolated as a cosmid clone and sequenced. The 5′ and 3′ ends of the gene were determined by comparison with the AHA10 cDNA sequence. AHA10 is the most divergent isoform characterized in the Arabidopsis family. AHA10 appears to be expressed primarily in developing seeds, as indicated by Northern blot analysis of AHA10 mRNA and by the analysis of transgenic plants expressing a β-glucuronidase (GUS) reporter gene fused to an AHA10 promoter. Our results indicate that one function of this unusually large H+-ATPase gene family is to allow for expression of different isoforms in different cell types.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Boutry M, Michelet B, Goffeau A (1989) Molecular cloning of a family of plant genes encoding a protein homologous to plasma membrane H+-translocating ATPases. Biochem Biophys Res Commun 162:567–574
Briskin DP (1990) The plasma membrane H+-ATPase of higher plant cells: biochemistry and transport function. Biochem Biophys Acta 1019:95–109
DeWitt ND, Harper JF, Sussman MR (1991) Evidence for a plasma membrane proton pump in phloem cells of higher plants. The Plant J 1:121–128
Ewing NN, Wimmers LE, Meyer DJ, Chetelat RT, Bennett AB (1990) Molecular cloning of tomato plasma membrane H+-ATPase. Plant Physiol 94:1874–1881
Felle H (1982) Effects of fusicoccin upon membrane potential, resistance and current-voltage characteristics in root hairs of Sinapsis alba. Plant Sci Lett 25:219–225
Fordham-Skelton AP, Yarwood A, Croy RRD (1990) Synthesis of saporin gene probes from partial protein sequence data: use of inosine-oligonucleotides, genomic DNA and the polymerase chain reaction. Mol Gen Genet 221:134–138
Frohman MA, Dush MK, Martin GR (1988) Rapid production of full-length cDNAs from rate transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci USA 85:8998–9002
Ghislain M, Goffeau A (1991) The pma1 and pma2 H+-ATPases from Schizosaccharomyces pombe are functionally interchangeable. J Biol Chem 266:18276–18279
Girgis SI, Alevizaki M, Denny P, Ferrier GJM, Legon S (1988) Generation of DNA probes for peptides with highly degenerate codons using mixed primer PCR. Nucleic Acids Res 16:10371
Goffeau A, Slayman CW (1981) The proton-translocating ATPase of the fungal plasma membrane. Biochem Biophys Acta 639:197–223
Habener JF, Vo CD, Le DB, Gryan GP, Ercolani L, Wang AHJ (1988) 5-Fluorodeoxyuridine as an alternative to the synthesis of mixed hybridization probes for the detection of specific gene sequences. Proc Natl Acad Sci USA 85:1735–1739
Harper JF, Huson KS, Kirk DL (1987) Use of repetitive sequences to identify DNA polymorphisms linked to regA, a developmentally important locus in Voluox. Genes Dev 1:573–584
Harper JF, Surowy TK, Sussman MR (1989) Molecular cloning and sequence of cDNA encoding the plasma membrane proton pump of Arabidopsis thaliana. Proc Natl Acad Sci USA 86:1234–1238
Harper IF, Manney L, DeWitt ND, Yoo MH, Sussman MR (1990) The Arabidopsis thaliana plasma membrane H+-ATPase multigene family. J Biol Chem 265:13601–13608
Houline G, Boutry M (1994) Identification of an Arabidopsis thaliana gene encoding a plasma membrane H+-ATPase whose expression is restricted to another tissue. Plant J 5:311–317
Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405
Johansson F, Sommarin M, Larsson C (1993) Fusicoccin activates the plasma membrane H+-ATPase by a mechanism involving the C-terminal inhibitory domain. The Plant Cell 5:321–327
Knoth K, Roberds S, Potect C, Tamkun M (1988) Highly degenerate, inosine-containing primers specifically amplify rare cDNA using the polymerise chain reaction. Nucleic Acids Res 16:10932
Kwok S, Kellog DE, McKinney N, Spasic D, Goda L, Levenson C, Sninsky JJ (1990) Effects of primer-template mismatches on the polymerase chain reaction: human immunodeficiency virus type 1 model studies. Nucleic Acids Res 18:999–1005
Lingrel JB, Orlowski J, Shull MM, Price EM (1990) Molecular genetics of Na, K-ATPase. Prog Nucleic Acid Res Mol Biol 38:37–89
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor New York
Moremen KW (1989) Isolation of a rat liver golgi mannosidase II clone by mixed oligonucleotide-primed amplification of cDNA. Proc Natl Acad Sci USA 86:5276–5280
Moriau L, Bogaerts P, Jonnizus J-L, Boutry M (1993) Identification and characterization of a second plasma membrane H+-ATPase gene subfamily in Nicotiana plumbaginifolia. Plant Mol Biol 21:955–963
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4231–4325
Offler CE, Nerlich SM, Patrick JW (1989) Pathway of photosynthate transfer in the developing seed of Vicia faba L. transfer in relation to seed anatomy. J Exp Bot 40:769–780
Olszewski NE, Martin FB, Ausubel FM (1988) Specialized binary vector for plant transformation: expression of the Arabidopsis thaliana AHAS gene in Nicotiana tabacum. Nucleic Acids Res 16:10765–10781
Palmgren MG, Christensen G (1993) Complementation in situ of the yeast plasma membrane H+-ATPase gene penal by an H+-ATPase gene from a heterologous species. FEBS Lett 317:216–222
Palmgren MG, Christensen G (1994) Functional comparisons between plant plasma membrane H+-ATPase isoforms expressed in yeast. J Biol Chem 269:3027–3033
Palmgren MG, Larsson C, Sommarin M (1990) Proteolytic activation of the plant plasma membrane H+-ATPase by removal of a terminal segment. J Biol Chem 265:13423–13426
Palmgren MG, Sommarin M, Serrano R, Larsson C (1991) Identification of an autoinhibitory domain in the C-terminal region of the plant plasma membrane H+-ATPase. J Biol Chem 266:20470–20475
Pardo JM, Serrano R (1989) Structure of a plasma membrane H+-ATPase gene from the plant Arabidopsis thaliana. J Biol Chem 264:8557–8562
Rajkumar VP, Dekker EE (1990) PCR amplification of an Escherichia coli gene using mixed primers containing deoxyinosine at ambiguous positions in degenerate amino acid codons. Nucleic Acids Res 18:3080
Rao R, Slayman CW (1992) Mutagenesis of the yeast plasma mem brane H+-ATPase. Biophys J 62:228–237
Rochester DE, Winter JA, Shah DM (1986) The structure and expression of maize genes encoding the major heat shock proteins hsp70. EMBO J 5:451–458
Schlesser A, Ulaszewski S, Ghiaslain M, Goffeau A (1988) A second trasport ATPase gene in Saccharomyces cerevisiae. J Biol Chem 263:19480–19487
Schroeder JI, Hedrich R (1989) Involvement of ion channels and active transport in osmoregulation and signaling of higher plant cells. Trends Biochem Sci 14:187–192
Schulz P, Jensen WA (1969) Capsella embryogenesis: the suspensor and the basal cell. Protoplasma 67:139–163
Schulz P, Jensen WA (1971) Capsella embryogenesis: the chalazal proliferating tissue. J Cell Sci 8:201–227
Schulz SP, Jensen WA (1974) Capsella embryogenesis: the development of the free nuclear endosperm. Protoplasma 80:183–205
Serrano R (1989) Structure and function of plasma membrane ATPase. Annu Rev Plant Physiol Plant Mol Biol 40:61–94
Sussman MR, Harper JF (1989) Molecular biology of the plasma membrane of higher plants. The Plant Cell 1:953–960
Tautz D, Renz M (1983) An optimized freeze-squeeze method for the recovery of DNA fragments from agarose gels. Anal Biochem 132:14–19
Valvekens D, Van Montagu M, Van Ligsebettens M (1988) Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci USA 85:5536–5540
Varadi A, Gilmore-Heber M, Benz EJ Jr (1989) Amplification of the phosphorylation site-ATP-binding site cDNA fragment of the Na+, K+-ATPase and the Ca2+-ATPase of Drosphila melanogaster by polymerase chain reaction. FEES Lett 258:203–207
Villalba JM, Lutzelschwab M, Serrano R (1991) Immunocytolocalization of plasma membrane H+-ATPase in maize coleoptiles and enclosed leaves. Planta 185:458–461
Wach A, Schlesser A, Goffeau A (1992) An alignment of 17 deduced protein sequences from plant, fungi, and ciliate H+-ATPase genes. J Bioenerg Biomembr 24:309–317
Wilks AF (1989) Two putative protein-tyrosine kinases identified by application of the polymerase chain reaction. Proc Natl Acad Sci USA 86:1603–1607
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Harper, J.F., Manney, L. & Sussman, M.R. The plasma membrane H+-ATPase gene family in Arabidopsis: genomic sequence of AHA10 which is expressed primarily in developing seeds. Molec. Gen. Genet. 244, 572–587 (1994). https://doi.org/10.1007/BF00282747
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00282747