Arabidopsis Tic62 and ferredoxin-NADP(H) oxidoreductase form light-regulated complexes that are integrated into the chloroplast redox poise.
Translocation of nuclear-encoded preproteins across the inner envelope of chloroplasts is catalyzed by the Tic translocon, consisting of Tic110, Tic40, Tic62, Tic55, Tic32, Tic20, and Tic22. Tic62 was proposed to act as a redox sensor of the complex because of its redox-dependent shuttling between envelope and stroma and its specific interaction with the photosynthetic protein ferredoxin-NADP(H) oxidoreductase (FNR). However, the nature of this close relationship so far remained enigmatic. A putative additional localization of Tic62 at the thylakoids mandated further studies examining how this feature might be involved in the respective redox sensing pathway and the interaction with its partner protein. Therefore, both the association with FNR and the physiological role of the third, thylakoid-bound pool of Tic62 were investigated in detail. Coexpression analysis indicates that Tic62 has similar expression patterns as genes involved in photosynthetic functions and protein turnover. At the thylakoids, Tic62 and FNR form high molecular weight complexes that are not involved in photosynthetic electron transfer but are dynamically regulated by light signals and the stromal pH. Structural analyses reveal that Tic62 binds to FNR in a novel binding mode for flavoproteins, with a major contribution from hydrophobic interactions. Moreover, in absence of Tic62, membrane binding and stability of FNR are drastically reduced. We conclude that Tic62 represents a major FNR interaction partner not only at the envelope and in the stroma, but also at the thylakoids of Arabidopsis thaliana and perhaps all flowering plants. Association with Tic62 stabilizes FNR and is involved in its dynamic and light-dependent membrane tethering.
Benz
J P
JP
Munich Center for Integrated Protein Science CiPS, Ludwig-Maximilians-Universität München, D-81377 Munich, Germany.
Stengel
A
A
Lintala
M
M
Lee
Y-H
YH
Weber
A
A
Philippar
K
K
Gügel
I L
IL
Kaieda
S
S
Ikegami
T
T
Mulo
P
P
Soll
J
J
Bölter
B
B
D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002736:Chloroplasts / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D055786:Gene Knockout Techniques; D008027:Light; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D016254:Mutagenesis, Insertional; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D010788:Photosynthesis; D011485:Protein Binding; D025941:Protein Interaction Mapping; D018749:RNA, Plant / Q000235:genetics
D029681:Arabidopsis Proteins; D004269:DNA, Bacterial; D026901:Membrane Transport Proteins; D018749:RNA, Plant; C036483:T-DNA; C520728:Tic62 protein, Arabidopsis; D010088:Oxidoreductases; C028878:ferredoxin-NAD+ reductase