cDNA and genomic cloning of sugar beet V-type H+- ATPase subunit A and c isoforms: evidence for coordinate expression during plant development and coordinate induction in response to high salinity
Identifieur interne : 003875 ( Main/Exploration ); précédent : 003874; suivant : 003876cDNA and genomic cloning of sugar beet V-type H+- ATPase subunit A and c isoforms: evidence for coordinate expression during plant development and coordinate induction in response to high salinity
Auteurs : Angelika Lehr [Allemagne] ; Matthias Kirsch [Allemagne] ; Ruth Viereck [Allemagne] ; Joachim Schiemann [Allemagne] ; Thomas Rausch [Allemagne]Source :
- Plant Molecular Biology [ 0167-4412 ] ; 1999-02-01.
English descriptors
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Abstract
Abstract: The plant V-type H+-ATPase (V-ATPase) does not only serve basic housekeeping functions but is also involved in stress- induced NaCl sequestration during salinity stress. To address the question whether the same isoforms conferring housekeeping functions are equally involved in the response to high salinity, we have isolated cDNA clones for subunits A and c, as representing the peripheral V1 complex and the membrane-integral V0 complex, respectively, from the halotolerant sugar beet (Beta vulgaris L., diploid variety). RNA blot analysis with gene-specific probes revealed a coordinate expression of the cloned subunit A and c isoforms during plant development and in response to high salinity. Also, in rapidly dividing suspension-cultured cells with 10-fold increased transcript amounts as compared to young leaf tissue, the ratio of transcripts for both genes was similar to the ratio found for transcripts in leaves of different age. We have then isolated partial genomic clones (BVA/70 for Beta V- ATPase 70 kDa subunit; BVA/16-1 for Beta V- ATPase 16 kDa subunit), including the promoter regions. Transcription start mapping revealed long 5′-UTR leader sequences (230 and 172 bases, respectively) for both genes. Both promoters contain putative G-box motifs in similar distance to the TATA boxes. For a quantitative comparison of relative promoter strength, the BVA/70 and BVA/16-1 promoters linked to the luciferase reporter gene (LUC) were delivered to sugar beet suspension-cultured cells by particle bombardment. The BVA/16-1 promoter showed a 1.7-fold higher activity as compared with the BVA/70 promoter. Salt treatment induced an increase of BVA/70 (+70%) and BVA/16-1 (+57%) promoter activities, concomitant with increased transcript amounts. The following sequences have been deposited at the EMBL database X98767: Beta vulgaris V-ATPase subunit A, cDNA clone; X98851, B. vulgaris V-ATPase subunit c isoform 1, cDNA clone; Y11038, B. vulgaris V-ATPase subunit A, partial genomic clone; Y11037, B. vulgaris V-ATPase subunit c isoform 1, partial genomic clone.
Url:
DOI: 10.1023/A:1006158310891
Affiliations:
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Genetics and Biochemistry</title><title level="j" type="abbrev">Plant Mol Biol</title><idno type="ISSN">0167-4412</idno><idno type="eISSN">1573-5028</idno><imprint><publisher>Kluwer Academic Publishers</publisher><pubPlace>Dordrecht</pubPlace><date type="published" when="1999-02-01">1999-02-01</date><biblScope unit="volume">39</biblScope><biblScope unit="issue">3</biblScope><biblScope unit="page" from="463">463</biblScope><biblScope unit="page" to="475">475</biblScope></imprint><idno type="ISSN">0167-4412</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0167-4412</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Beta vulgaris L. (sugar beet)</term><term>V-ATPase</term><term>coordinate expression</term><term>isoforms</term><term>promoter structure</term><term>subunit A</term><term>subunit c</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: The plant V-type H+-ATPase (V-ATPase) does not only serve basic housekeeping functions but is also involved in stress- induced NaCl sequestration during salinity stress. To address the question whether the same isoforms conferring housekeeping functions are equally involved in the response to high salinity, we have isolated cDNA clones for subunits A and c, as representing the peripheral V1 complex and the membrane-integral V0 complex, respectively, from the halotolerant sugar beet (Beta vulgaris L., diploid variety). RNA blot analysis with gene-specific probes revealed a coordinate expression of the cloned subunit A and c isoforms during plant development and in response to high salinity. Also, in rapidly dividing suspension-cultured cells with 10-fold increased transcript amounts as compared to young leaf tissue, the ratio of transcripts for both genes was similar to the ratio found for transcripts in leaves of different age. We have then isolated partial genomic clones (BVA/70 for Beta V- ATPase 70 kDa subunit; BVA/16-1 for Beta V- ATPase 16 kDa subunit), including the promoter regions. Transcription start mapping revealed long 5′-UTR leader sequences (230 and 172 bases, respectively) for both genes. Both promoters contain putative G-box motifs in similar distance to the TATA boxes. For a quantitative comparison of relative promoter strength, the BVA/70 and BVA/16-1 promoters linked to the luciferase reporter gene (LUC) were delivered to sugar beet suspension-cultured cells by particle bombardment. The BVA/16-1 promoter showed a 1.7-fold higher activity as compared with the BVA/70 promoter. Salt treatment induced an increase of BVA/70 (+70%) and BVA/16-1 (+57%) promoter activities, concomitant with increased transcript amounts. The following sequences have been deposited at the EMBL database X98767: Beta vulgaris V-ATPase subunit A, cDNA clone; X98851, B. vulgaris V-ATPase subunit c isoform 1, cDNA clone; Y11038, B. vulgaris V-ATPase subunit A, partial genomic clone; Y11037, B. vulgaris V-ATPase subunit c isoform 1, partial genomic clone.</div></front></TEI><affiliations><list><country><li>Allemagne</li></country><region><li>Bade-Wurtemberg</li><li>District de Karlsruhe</li></region><settlement><li>Heidelberg</li></settlement></list><tree><country name="Allemagne"><region name="Bade-Wurtemberg"><name sortKey="Lehr, Angelika" sort="Lehr, Angelika" uniqKey="Lehr A" first="Angelika" last="Lehr">Angelika Lehr</name></region><name sortKey="Kirsch, Matthias" sort="Kirsch, Matthias" uniqKey="Kirsch M" first="Matthias" last="Kirsch">Matthias Kirsch</name><name sortKey="Rausch, Thomas" sort="Rausch, Thomas" uniqKey="Rausch T" first="Thomas" last="Rausch">Thomas Rausch</name><name sortKey="Schiemann, Joachim" sort="Schiemann, Joachim" uniqKey="Schiemann J" first="Joachim" last="Schiemann">Joachim Schiemann</name><name sortKey="Viereck, Ruth" sort="Viereck, Ruth" uniqKey="Viereck R" first="Ruth" last="Viereck">Ruth Viereck</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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