Isolation and characterisation of P-type H+-ATPase gene from potato

ArticleinPlant Molecular Biology 26(3):979-88 · December 1994with31 Reads
DOI: 10.1007/BF00028864 · Source: PubMed
H(+)-ATPase cDNAs were identified in a potato leaf library using an Arabidopsis gene as a probe. Based on their sequences, the clones could be grouped into at least two classes. A similar classification was obtained from the analysis of sequence data from four tobacco genes. Both potato genes are expressed in all tissues analysed, higher levels of expression were found in leaves and stem than in roots and tubers. For both genes, no significant differences in level of expression could be detected under a variety of conditions such as cold treatment, anaerobiosis, sucrose induction or treatment with a synthetic cytokinin. Only 2,4-D and prolonged periods of darkness lead to a slight reduction in mRNA levels. The reduction in darkness was compensated after transfer of the plants back into the light. Expression of the ATPase genes remained constant in transgenic plants which are inhibited in phloem loading due to antisense inhibition of the sucrose transporter. On the other hand, expression of the sucrose transporter is inducible by auxin and cytokinin but not by sucrose. Taken together, these data suggest that at least the two plasma membrane H(+)-ATPase genes analysed are rather constant in their expression and that either other genes respond to external stimuli or that most of the regulation occurs at the posttranscriptional level.
    • "The PM H + -ATPase enzyme is encoded by a multigene family (Portillo, 2000; Arango et al., 2003). Many studies have shown changes of in gene expression of the PM H + - ATPase in response to a variety of environmental factors, including salt stress (Binzel, 1995; Janicka-Russak and K1obus, 2007), dehydration (Surowy and Boyer, 1991), light conditions (Harms et al., 1994), mechanical stress (Oufattole et al., 2000), and low temperature (Janicka- Russak et al., 2011). It is known that, besides the genetic regulation of the proton pump, its activity might be fast modulated post-translationally at the protein level, mainly through reversible phosphorylation (Schaller and Sussman, 1988; Portillo, 2000). "
    [Show abstract] [Hide abstract] ABSTRACT: The effect of heavy metals on plasma membrane (PM) H+-ATPase (EC activity in cucumber (Cucumis sativus) roots was studied. The aim of this work was to explain the mechanism of modification of the PM H+-ATPase activity in plants subjected to heavy metals. Plants were treated with 10 μM Cd or Cu for 6 d. After 3 d exposure to the heavy metals, some of the plants were transferred to control conditions for a further 3 d (3/3 plants). The activity of PM H+-ATPase was found to be increased in plants treated with heavy metals. The highest activity measured as proton transport was observed in 3/3 plants. Estimation of transcript levels of C. sativus PM H+-ATPase in roots indicated that the action of Cd, but not Cu, affected the gene expression level. Transcript levels of C. sativus PM H+-ATPase (CsHA2, CsHA3, CsHA4, CsHA8, and CsHA9) genes increased in roots treated with Cd. Moreover, Western blot analysis with antibody against phosphothreonine and 14-3-3 protein indicated that increased activity of PM H+-ATPase under heavy-metal stress resulted from phosphorylation of the enzyme. It was found that Cu markedly increased the activity of catalase and ascorbate peroxidase and reduced the level of H2O2 in cucumber roots. In contrast, Cd did not affect these parameters. These results indicate that Cd and Cu can, in different ways, lead to modification of PM H+-ATPase activity. Additionally, it was observed that treatment of plants with heavy metals led to an increased level of heat-shock proteins in the tissues. This suggests that the plants had started adaptive processes to survive adverse conditions, and increased PM H+-ATPase activity could further enhance the repair processes in heavy-metal-stressed plants.
    Full-text · Article · Mar 2012
    • "Only in the case of SUT2 from tomato plants was a positive effect on sucrose transporter expression detected (Barker et al. 2000). The accumulation of the SUT1 mRNA from potato is inducible by auxin and cytokinin (Harms et al. 1994), and this up-regulation is also reflected at the protein level (He et al. 2008). Analysis of the genomic StSUT1 sequence revealed the presence of a putative-binding domain of the auxin response factor, ARF (auxin responsive element AuxRE), in the third intron. "
    [Show abstract] [Hide abstract] ABSTRACT: Sucrose transporters are essential proteins for the allocation of assimilates from source to sink. With increasing information about their function and the role of the sucrose molecule, acting as an informational and signaling molecule, new questions arise on how sucrose transporters could interact or coordinate metabolic pathways and developmental processes and to integrate whole plant communication. New information about the phloem mobility of sucrose transporter mRNAs and other phloem mobile signals is also available, shedding light on the long-distance transport of leaf-derived information to terminal sink organs. Recent advances on the subcellular localisation and function of sucrose transporters, sucrose facilitators and sucrose transporter-like proteins open new questions about the role of membrane compartmentation on the dimerization, endocytosis, degradation and signaling of plant sucrose transporters.
    Full-text · Article · Jan 2011 · Planta
    • "This hypothesis is supported by the fact that at least one PM H ? -ATPase isoform expression also increases during tuber development, although more studies are necessary to corroborate this idea, especially considering the differences in the expression profile and in the magnitude of induction. The primers used to determine the expression of potato PM H ? -ATPase would detect the PHA2 and PHA1 isoforms, which are the only PM H ? -ATPases reported for S. tuberosum (Harms et al. 1994), however, the possibility that other isoforms may be amplified by the primers cannot be excluded. It would be interesting in the future to identify and characterize the isoforms that are induced during tuber development. "
    [Show abstract] [Hide abstract] ABSTRACT: Plasma membrane proton pumps (PM H(+)-ATPases) are involved in several physiological processes, such as growth and development, and abiotic stress responses. The major regulators of the PM H(+)-ATPases are proteins of the 14-3-3 family, which stimulate its activity. In addition, a novel interaction partner of the AHA1 PM H(+)-ATPase, named PPI1 (proton pump interactor, isoform 1), was identified in Arabidopsis thaliana. This protein stimulates the activity of the proton pump in vitro. In this work, we report the characterization of an A. thaliana PPI1 homolog in potato (Solanum tuberosum L.) named StPPI1. The full-length coding sequence of StPPI1 was obtained. The open reading frame (ORF) encodes a protein of 629 amino acids showing 50% identity with A. thaliana PPI1 protein. The StPPI1 ORF is divided into seven exons split by six introns. Southern blot analysis suggests that StPPI1 belongs to a family of related genes. Recombinant StPPI1 stimulates H(+)-ATPase activity in vitro. Basal levels of StPPI1 transcripts are observed in all tissues, however, StPPI1 expression is higher in proliferative regions (shoot apex and flower buds), flowers and leaves than in shoots and roots. StPPI1 mRNA levels significantly increase during tuber development. StPPI1 is induced by salt stress and cold. Drought and mechanical wounding slightly increase StPPI1 transcript levels. In addition, the expression of SlPPI1, the tomato homolog of StPPI1, was determined under adverse environmental conditions in tomato plants. SlPPI1 mRNA levels are increased by drought and cold, but are unaffected by salt stress. Mechanical wounding slightly increases SlPPI1 expression.
    Full-text · Article · Dec 2010
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