Perez C, Michelet B, Ferrant V, Bogaerts P, Boutry M
Unité de Biochimie Physiologique, Université Catholique de Louvain, Louvain-la-Neuve, Belgium.
J Biol Chem. 1992 Jan 15;267(2):1204-11.
Genomic and cDNA clones for the three members of a gene subfamily (pma) encoding a plasma membrane H(+)-translocating ATPase in Nicotiana plumbaginifolia were isolated and sequenced. They are between 95 and 96% identical at the deduced amino acid sequence level. Sequence comparisons with the corresponding tomato genes (Ewing, N.N., Wimmers, L.E., Meyer, D.J., Chetelat, R.T., and Bennett, A.B. (1990) Plant Physiol. 94, 1874-1881) indicate that divergence among the three N. plumbaginifolia pma genes occurred before the development of the Solanaceae family. Here, determination of pma1 transcription initiation sites reveals several 5' boundaries located 266 to 120 nucleotides upstream from the plasma membrane H(+)-ATPase translation initiation codon. The 5'-untranslated region contains a small open reading frame, 9 residues long. pma3 has a single, 264-nucleotide long 5' leader containing a 5-residue open reading frame. The latter is completely conserved in a corresponding tomato gene. These features suggest the possibility of translational regulation of plant pma genes. S1 nuclease protection assays on total cellular RNA isolated from different organs reveals that all three genes are expressed in leaf, stem, flower, and root tissues, albeit at different levels according to the organ and gene. The different genes for the plant H(+)-translocating ATPase are thus subject to differential regulation of transcription, possibly related to specific aspects of enzyme function.
分离并测序了烟草中编码质膜H⁺转运ATPase的一个基因亚家族(pma)的三个成员的基因组和cDNA克隆。它们在推导的氨基酸序列水平上有95%到96%的同一性。与相应番茄基因(Ewing, N.N., Wimmers, L.E., Meyer, D.J., Chetelat, R.T., and Bennett, A.B. (1990) Plant Physiol. 94, 1874 - 1881)的序列比较表明,烟草的三个pma基因之间的分歧发生在茄科植物进化之前。在这里,pma1转录起始位点的确定揭示了几个位于质膜H⁺ - ATPase翻译起始密码子上游266至120个核苷酸处的5'边界。5'非翻译区包含一个9个残基长的小开放阅读框。pma3有一个单一的、264个核苷酸长的5'前导序列,包含一个5个残基的开放阅读框。后者在相应的番茄基因中完全保守。这些特征表明植物pma基因存在翻译调控的可能性。对从不同器官分离的总细胞RNA进行的S1核酸酶保护分析表明,所有三个基因在叶、茎、花和根组织中均有表达,尽管根据器官和基因的不同表达水平有所差异。因此,植物H⁺转运ATPase的不同基因受到转录的差异调控,这可能与酶功能的特定方面有关。