A stromal region of cytochrome b6f subunit IV is involved in the activation of the Stt7 kinase in Chlamydomonas

Proc Natl Acad Sci U S A. 2017 Nov 7;114(45):12063-12068. doi: 10.1073/pnas.1713343114. Epub 2017 Oct 24.

Abstract

The cytochrome (cyt) b6f complex 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 cyt b6f activates the Stt7 kinase through a mechanism that is still poorly understood. After random mutagenesis of the chloroplast petD gene, coding for subunit IV of the cyt b6f complex, and complementation of a ΔpetD host 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 cyt b6f subunit IV are crucial for state transitions. In vitro reconstitution experiments with purified cyt b6f and recombinant Stt7 kinase domain show that cyt b6f enhances Stt7 autophosphorylation and that the Arg125 residue is directly involved in this process. The peripheral stromal structure of the cyt b6f complex had, until now, no reported function. Evidence is now provided of a direct interaction with Stt7 on the stromal side of the membrane.

Keywords: Stt7 kinase; cytochrome b6f complex; random mutagenesis; state transitions; structure–function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chlamydomonas / metabolism*
  • Chlorophyll / metabolism
  • Chloroplasts / metabolism
  • Cytochrome b6f Complex / metabolism*
  • Light-Harvesting Protein Complexes / metabolism
  • Oxidation-Reduction
  • Phosphorylation / physiology
  • Photosystem I Protein Complex / metabolism
  • Photosystem II Protein Complex / metabolism
  • Plastoquinone / metabolism
  • Protein Kinases / metabolism*

Substances

  • Light-Harvesting Protein Complexes
  • Photosystem I Protein Complex
  • Photosystem II Protein Complex
  • Chlorophyll
  • Cytochrome b6f Complex
  • Protein Kinases
  • Plastoquinone