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Plant Physiol, November 2001, Vol. 127, pp. 1044-1052
Chloroplast Transcription at Different Light Intensities.
Glutathione-Mediated Phosphorylation of the Major RNA Polymerase
Involved in Redox-Regulated Organellar Gene
Expression1
Elena
Baena-González,2
Sacha
Baginsky,23
Paula
Mulo,
Holger
Summer,
Eva-Mari
Aro, and
Gerhard
Link*
Department of Biology, University of Turku, FIN-20014 Turku,
Finland (E.B.-G., P.M., E.-M.A.); and Plant Cell Physiology,
University of Bochum, D-44780 Bochum, Germany (S.B., H.S.,
G.L.)
Previous studies using purified RNA polymerase from mustard
(Sinapis alba) chloroplasts showed control of
transcription by an associated protein kinase. This kinase was found to
respond to reversible thiol/disulfide formation mediated by glutathione (GSH), although at concentrations exceeding those thought to exist in
vivo. In the present study, several lines of evidence are presented to
substantiate the functioning of this regulation mechanism, also in
vivo: (a) Studies on the polymerase-associated transcription kinase
revealed that at appropriate ATP levels, GSH concentrations similar to
those in vivo are sufficient to modulate the kinase activity; (b) GSH
measurements from isolated mustard chloroplasts showed considerable
differences in response to light intensity; (c) this was reflected by
run-on transcription rates in isolated chloroplasts that were generally
higher if organelles were prepared from seedlings incubated under
high-light as compared with growth-light conditions; (d) the notion of
a general transcriptional switch was strengthened by in vitro
experiments showing that the kinase not only affects the transcription
of a photosynthetic gene (psbA) but also that of a
non-photosynthetic gene (trnQ); and (e) the polymerase-kinase complex revealed specific differences in the phosphorylation state of polypeptides depending on the light intensity to which the seedlings had been exposed prior to chloroplast isolation. Taken together, these data are consistent with GSH and
phosphorylation-dependent regulation of chloroplast transcription in vivo.
1
This work was supported by the Academy of
Finland (grant to E.-M.A.), by the Deutsche Akademische Austausdienst,
by the Deutsche Forschungsgemeinschaft, and by the Fonds der Chemischen
Industrie (to G.L.).
2
These authors contributed equally to the paper.
3
Present address: Plant Science Institute, Swiss Federal
Institute of Technology, ETH-Zentrum, CH-8092 Zürich, Switzerland.
*
Corresponding author; e-mail
Gerhard.Link{at}ruhr-uni-bochum.de; fax 49-234-3214-188.
© 2001 American Society of Plant Physiologists
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