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Plant Physiol, February 2001, Vol. 125, pp. 969-981
Redox Control of ntcA Gene Expression
in Synechocystis sp. PCC 6803. Nitrogen Availability
and Electron Transport Regulate the Levels of the NtcA
Protein1
Miguel
Alfonso,2*
Irène
Perewoska, and
Diana
Kirilovsky3
Unité Mixte de Recherche, 8543 Photorégulation et
Dynamique des Membranes Végetales, Centre National de la
Recherche Scientifique, Ecole Normale Supériéure, 46 rue
d'Ulm, 75230 Paris cedex 05, France
In this work we have studied the influence of the cellular redox
status in the expression of the Synechocystis sp.
PCC 6803 ntcA gene. Two different ntcA
transcripts with different 5' ends were detected, depending on the
different dark/light or nitrogen availability conditions. Accumulation
of a 0.8-kb ntcA message was light and nitrogen
dependent, whereas a longer 1.2-kb ntcA transcript was
neither light nor nitrogen regulated. NtcA protein levels increased
concomitantly with the accumulation of the 0.8-kb ntcA
transcript. The light-dependent accumulation of the ntcA gene and the NtcA protein was sensitive to electron transport inhibitors. In addition, Glc-grown Synechocystis sp.
cells showed a similar ntcA expression pattern in
darkness to that observed under illumination. These data suggested that
electron transport, and not light per se may regulate
ntcA gene expression. Primer extension analysis,
together with gel mobility-shift assays, demonstrated that in vitro,
the Synechocystis sp. NtcA protein specifically bound to
the putative promoter region from the light/nitrogen-dependent ntcA transcript but not to that from the constitutive
1.2-kb ntcA mRNA. Band-shift experiments carried out in
the presence of thiol oxidizing/modifiying agents and different
reducing/oxidizing conditions suggested that NtcA binding to its own
promoter was under a thiol-dependent redox mechanism. Our results
suggest that the cellular redox status plays a central role in the
autoregulatory mechanism of the NtcA protein.
1
This work was supported by grants from the
Spanish Ministry of Education (no. PB98-1632 to M.A.) and the Centre
National de la Recherche Scientifique (to D.K.). M.A. is recipient of a
Postdoctoral Contract from Ministerio de Educación y Cultura.
2
Present address: Departamento de Nutrición
Vegetal, Estación Experimental de Aula Dei, Apartado 202, 50080 Zaragoza, Spain.
3
Present address: Section de Bioénergétique,
Département de Biologie Cellulaire et Moléculaire,
Bât. 532, CEA Saclay, 91191 Gif-sur-Yvette, France.
*
Corresponding author; e-mail alfonso{at}eead.csic.es; fax
34-76-575620.
© 2001 American Society of Plant Physiologists
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