Plant Physiology Preview
Published on April 28, 2006; 10.1104/pp.106.080150
Received March 14, 2006
Returned for revision April 18, 2006
Accepted April 21, 2006
High light response of the thylakoid proteome in Arabidopsis thaliana wild type and the ascorbate deficient mutant vtc2-2; a comparative proteomics study
Lisa Giacomelli , Andrea Rudella , and Klaas Jan van Wijk *
Department of Plant Biology, Cornell University, Ithaca, New York 14853
* Corresponding author; email: kv35{at}cornell.edu.
The thylakoid proteome of chloroplasts contains multiple proteins involved in anti-oxidative defense, protein folding and repair. To understand this functional protein network, we analyzed the quantitative response of the thylakoid-associated proteome of Arabidopsis wt and the ascorbate deficient mutant vtc2-2 after transition to high light (1000 µmol photons.m-2.s-1). The soluble thylakoid proteomes of wt and vtc2-2 were compared after 0, 1, 3 and 5 days of high light using 2-dimensional gels with three independent experiments, followed by a multi-variant statistical analysis and tandem mass spectrometry. After 5 days of high light, both wt and vtc2-2 plants accumulated anthocyanins, increased their total ascorbate content and lost 10% of Photosystem II efficiency, but showed no bleaching. Anthocyanin and total ascorbate concentrations in vtc2-2 were respectively 34% and 20% of wt, potentially leading to enhanced oxidative stress in vtc2-2. 45 proteins spots significantly changed as a consequence of genotype, light treatment or both. Independent confirmation was obtained from western blots. The most significant response was the up-regulation of thylakoid YCF37 likely involved in Photosystem I assembly and specific fibrillins, a flavin reductase-like protein and an aldolase, each located in thylakoid associated plastoglobules. Fe-SOD was down regulated in vtc2-2, while Cu,Zn-SOD was up-regulated. vtc2-2 also showed a systematic up-regulation of a steroid dehydrogenase-like protein. A number of other stress related proteins, several thylakoid proteases and lumenal isomerases did not change, while PsbS increased in wt upon light stress. These findings are discussed in terms of plastid metabolism and oxidative stress defense, and emphasize that understanding of the chloroplast stress response network must include the enzymatic role of plastoglobules.
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