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cDNA-Array
Redox regulation of
metabolic reactions and gene expression is a common principle in
adjusting photosynthetically active cells to changing environmental
conditions, for example to balance light absorption to energy
consumption. Various levels of redox controlled adaptations can be
distinguished and range from fast metabolic responses such as state
transition to slow changes of structure which involve alterations in
gene expression.
Three specific cDNA
arrays were developed to quantify the transcript levels of nuclear and
plastid genes involved in redox regulation. One array (Redox-Array) contains candidate
genes encoding antioxidative and redox-associated enzymes, redox
modulators and stress inducible proteins. The second array comprises all
members of nuclear encoded gene families like peroxiredoxins,
cyclophilins, glutaredoxins, thioredoxins, catalases, superoxide
dismutases, ascorbate peroxidases and ascorbate oxidases (Redoxin-Array). The third array
contains all chloroplast genes encoding proteins (Plastome-Array). The sets of DNA
sequences were amplified via RT-PCR and spotted on Nylon membranes. The
sensitivity and reliability of the experimental system was evaluated and
a procedure was developed for detecting differential gene expression at
the transcript level. At the moment, a fourth array containing all
mitochondrial genes encoding proteins (Chondriome-Array) is in preparation.
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The
establishment of varying photosynthesis conditions (low or high CO2,
low or high O2, high or low light) in combination with
different effectors causes changes in the electron pressure and affects
the transcript profile monitored by macroarrays.Comparing these results
of different conditions as well as the response to redox active
effectors and different growth conditions and the use of metabolic and
redox disturbed mutants can lead to a better understanding of the redox
regulation of gene expression, the coordination of chloroplast,
mitochondrial and nuclear gene expression and may further narrow down
the nature of possible redox signals. |
Overview over cDNA-Arrays:
Redox-Array
antioxidative
enzymes, redox-associated enzymes, redox modulators, transcription
factors and modulators, enzymes of the primary metabolism, light
harvesting complexes, PR-proteins, diamide-inducible genes
Plastom-Array
ATPase
-subunits, cytochrome, subunits of PSI and PSII, RubisCo-LSU, subunits
of NADH-dehydrogenase, ribosomal proteins, RNA-polymerase-subunits,
ycfs, maturase
Redoxin-Array
peroxiredoxins,
cyclophilins, glutaredoxins, catalases, superoxide dismutases,
ascorbate peroxidases, ascorbate oxidases, glutathione reductases,
glutathione synthases, thioredoxins
Chondriom-Array
ribosomal
proteins, subunits of NADH-dehydrogenase, subunits of ATPase, cytochrom
c-oxidase, apocytochroms, 52 orfs, alternative oxidase, glycolate
oxidase, glycine decarboxylase and nuclear genes encoding various
mitochondrial proteins
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cDNA-Array:
exemplary hybridisation pattern and organisation
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Literature
Becker B., Holtgrefe S., Jung S., Wunrau C.,
Kandlbinder A., Baier M., Dietz K.J. (2006) Influence of the photoperiod on redox
regulation and stress responses in Arabidopsis thaliana L. (Heynh.) plants
under long- and short-day conditions. Planta 224, 380-393
Lamkemeyer P., Laxa M., Collin V., Li W., Finkemeier I.,
Schöttler M.A., Holtkamp V., Tognetti V.B., Issakidis-Bourguet E., Kandlbinder
A., Weis E., Miginiac-Maslow M., Dietz, K.J. (2006) Peroxiredoxin Q of Arabidopsis thaliana
is attached to the thylakoids and functions in context of photosynthesis. Plant
Journal 45, 968-981
Pfalz J., Liere K., Kandlbinder A., Dietz K.J.,
Oelmueller R. (2006)
PTAC2, -6, and -12 are components of the transcriptionally active plastid
chromosome that are required for plastid gene expression. Plant Cell 18,
176-197
Finkemeier I., Goodman M., Lamkemeyer
P., Kandlbinder A., Sweetlove L.J.,
Dietz, K.J. (2005)
The mitochondrial type II peroxiredoxin F is essential for redox homeostasis
and root growth of Arabidopsis thaliana under stress. JBC 280, 12168-12180
Kandlbinder A., Finkemeier I.,
Wormuth D., Hanitsch M., Dietz K.J. (2003) The antioxidant status of photosynthesising
leaves under nutrient deficiency: redox regulation, gene expression and
antioxidant activity in Arabidopsis thaliana. Physiologia Plantarum, Special
issue on Redox Regulation, (120), 63-74
This work is supported by the German Research Foundation (DFG) within
the collaboration network of Forschergruppe 387 (TP3).
Contact:Andrea Kandlbinder
(W5-120)
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