A stromal region of cytochromeb6fsubunit IV is involved in the activation of the Stt7 kinase inChlamydomonas

Abstract

The cytochrome (cyt)b6fcomplex and Stt7 kinase regulate the antenna sizes of photosystems I and II through state transitions, which are mediated by a reversible phosphorylation of light harvesting complexes II, depending on the redox state of the plastoquinone pool. When the pool is reduced, the cytb6factivates the Stt7 kinase through a mechanism that is still poorly understood. After random mutagenesis of the chloroplastpetDgene, coding for subunit IV of the cytb6fcomplex, and complementation of a ΔpetDhost strain by chloroplast transformation, we screened for impaired state transitions in vivo by chlorophyll fluorescence imaging. We show that residues Asn122, Tyr124, and Arg125 in the stromal loop linking helices F and G of cytb6fsubunit IV are crucial for state transitions. In vitro reconstitution experiments with purified cytb6fand recombinant Stt7 kinase domain show that cytb6fenhances Stt7 autophosphorylation and that the Arg125 residue is directly involved in this process. The peripheral stromal structure of the cytb6fcomplex had, until now, no reported function. Evidence is now provided of a direct interaction with Stt7 on the stromal side of the membrane.