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Global gene expression under nitrogen starvation in Xylella fastidiosa: contribution of the σ54 regulon

Identifieur interne : 000C20 ( Ncbi/Checkpoint ); précédent : 000C19; suivant : 000C21

Global gene expression under nitrogen starvation in Xylella fastidiosa: contribution of the σ54 regulon

Auteurs : José F. Da Silva Neto [Brésil] ; Tie Koide [Brésil] ; Suely L. Gomes [Brésil] ; Marilis V. Marques [Brésil]

Source :

RBID : PMC:3224663

Abstract

Background

Xylella fastidiosa, a Gram-negative fastidious bacterium, grows in the xylem of several plants causing diseases such as citrus variegated chlorosis. As the xylem sap contains low concentrations of amino acids and other compounds, X. fastidiosa needs to cope with nitrogen limitation in its natural habitat.

Results

In this work, we performed a whole-genome microarray analysis of the X. fastidiosa nitrogen starvation response. A time course experiment (2, 8 and 12 hours) of cultures grown in defined medium under nitrogen starvation revealed many differentially expressed genes, such as those related to transport, nitrogen assimilation, amino acid biosynthesis, transcriptional regulation, and many genes encoding hypothetical proteins. In addition, a decrease in the expression levels of many genes involved in carbon metabolism and energy generation pathways was also observed. Comparison of gene expression profiles between the wild type strain and the rpoN null mutant allowed the identification of genes directly or indirectly induced by nitrogen starvation in a σ54-dependent manner. A more complete picture of the σ54 regulon was achieved by combining the transcriptome data with an in silico search for potential σ54-dependent promoters, using a position weight matrix approach. One of these σ54-predicted binding sites, located upstream of the glnA gene (encoding glutamine synthetase), was validated by primer extension assays, confirming that this gene has a σ54-dependent promoter.

Conclusions

Together, these results show that nitrogen starvation causes intense changes in the X. fastidiosa transcriptome and some of these differentially expressed genes belong to the σ54 regulon.


Url:
DOI: 10.1186/1471-2180-10-231
PubMed: 20799976
PubMed Central: 3224663


Affiliations:


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PMC:3224663

Le document en format XML

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<title>Background</title>
<p>
<italic>Xylella fastidiosa</italic>
, a Gram-negative fastidious bacterium, grows in the xylem of several plants causing diseases such as citrus variegated chlorosis. As the xylem sap contains low concentrations of amino acids and other compounds,
<italic>X. fastidiosa </italic>
needs to cope with nitrogen limitation in its natural habitat.</p>
</sec>
<sec>
<title>Results</title>
<p>In this work, we performed a whole-genome microarray analysis of the
<italic>X. fastidiosa </italic>
nitrogen starvation response. A time course experiment (2, 8 and 12 hours) of cultures grown in defined medium under nitrogen starvation revealed many differentially expressed genes, such as those related to transport, nitrogen assimilation, amino acid biosynthesis, transcriptional regulation, and many genes encoding hypothetical proteins. In addition, a decrease in the expression levels of many genes involved in carbon metabolism and energy generation pathways was also observed. Comparison of gene expression profiles between the wild type strain and the
<italic>rpoN </italic>
null mutant allowed the identification of genes directly or indirectly induced by nitrogen starvation in a σ
<sup>54</sup>
-dependent manner. A more complete picture of the σ
<sup>54 </sup>
regulon was achieved by combining the transcriptome data with an
<italic>in silico </italic>
search for potential σ
<sup>54</sup>
-dependent promoters, using a position weight matrix approach. One of these σ
<sup>54</sup>
-predicted binding sites, located upstream of the
<italic>glnA </italic>
gene (encoding glutamine synthetase), was validated by primer extension assays, confirming that this gene has a σ
<sup>54</sup>
-dependent promoter.</p>
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<sec>
<title>Conclusions</title>
<p>Together, these results show that nitrogen starvation causes intense changes in the
<italic>X. fastidiosa </italic>
transcriptome and some of these differentially expressed genes belong to the σ
<sup>54 </sup>
regulon.</p>
</sec>
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<name sortKey="Krushkal, J" uniqKey="Krushkal J">J Krushkal</name>
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<author>
<name sortKey="Ueki, T" uniqKey="Ueki T">T Ueki</name>
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<author>
<name sortKey="Puljic, M" uniqKey="Puljic M">M Puljic</name>
</author>
<author>
<name sortKey="Sun, J" uniqKey="Sun J">J Sun</name>
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<author>
<name sortKey="Nu Ez, C" uniqKey="Nu Ez C">C Núñez</name>
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<author>
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<author>
<name sortKey="Didonato, R" uniqKey="Didonato R">R DiDonato</name>
</author>
<author>
<name sortKey="Postier, B" uniqKey="Postier B">B Postier</name>
</author>
<author>
<name sortKey="Adkins, Rm" uniqKey="Adkins R">RM Adkins</name>
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<author>
<name sortKey="Lovley, Dr" uniqKey="Lovley D">DR Lovley</name>
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<author>
<name sortKey="Pessi, G" uniqKey="Pessi G">G Pessi</name>
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<author>
<name sortKey="Weber, C" uniqKey="Weber C">C Weber</name>
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</author>
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</author>
<author>
<name sortKey="Fischer, Hm" uniqKey="Fischer H">HM Fischer</name>
</author>
<author>
<name sortKey="Hennecke, H" uniqKey="Hennecke H">H Hennecke</name>
</author>
</analytic>
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<analytic>
<author>
<name sortKey="Reitzer, Lj" uniqKey="Reitzer L">LJ Reitzer</name>
</author>
<author>
<name sortKey="Magasanik, B" uniqKey="Magasanik B">B Magasanik</name>
</author>
</analytic>
</biblStruct>
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<analytic>
<author>
<name sortKey="Craig, Nl" uniqKey="Craig N">NL Craig</name>
</author>
<author>
<name sortKey="Nash, Ha" uniqKey="Nash H">HA Nash</name>
</author>
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<biblStruct>
<analytic>
<author>
<name sortKey="Britto, Dt" uniqKey="Britto D">DT Britto</name>
</author>
<author>
<name sortKey="Siddiqi, My" uniqKey="Siddiqi M">MY Siddiqi</name>
</author>
<author>
<name sortKey="Glass, Adm" uniqKey="Glass A">ADM Glass</name>
</author>
<author>
<name sortKey="Kronzucker, Hj" uniqKey="Kronzucker H">HJ Kronzucker</name>
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<name sortKey="Schjoerring, Jk" uniqKey="Schjoerring J">JK Schjoerring</name>
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<author>
<name sortKey="Husted, S" uniqKey="Husted S">S Husted</name>
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<name sortKey="Mack, G" uniqKey="Mack G">G Mack</name>
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<author>
<name sortKey="Mattsson, M" uniqKey="Mattsson M">M Mattsson</name>
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</TEI>
<affiliations>
<list>
<country>
<li>Brésil</li>
</country>
<region>
<li>État de São Paulo</li>
</region>
<settlement>
<li>São Paulo</li>
</settlement>
<orgName>
<li>Université de São Paulo</li>
</orgName>
</list>
<tree>
<country name="Brésil">
<region name="État de São Paulo">
<name sortKey="Da Silva Neto, Jose F" sort="Da Silva Neto, Jose F" uniqKey="Da Silva Neto J" first="José F" last="Da Silva Neto">José F. Da Silva Neto</name>
</region>
<name sortKey="Gomes, Suely L" sort="Gomes, Suely L" uniqKey="Gomes S" first="Suely L" last="Gomes">Suely L. Gomes</name>
<name sortKey="Koide, Tie" sort="Koide, Tie" uniqKey="Koide T" first="Tie" last="Koide">Tie Koide</name>
<name sortKey="Koide, Tie" sort="Koide, Tie" uniqKey="Koide T" first="Tie" last="Koide">Tie Koide</name>
<name sortKey="Marques, Marilis V" sort="Marques, Marilis V" uniqKey="Marques M" first="Marilis V" last="Marques">Marilis V. Marques</name>
</country>
</tree>
</affiliations>
</record>

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