title: Novel thylakoid membrane GreenCut protein CPLD38 impacts accumulation of the cytochrome b6f complex and associated regulatory processes.
abstract: Based on previous comparative genomic analyses, a set of nearly 600 polypeptides was identified that is present in green algae and flowering and nonflowering plants but is not present (or is highly diverged) in nonphotosynthetic organisms. The gene encoding one of these "GreenCut" proteins, CPLD38, is in the same operon as ndhL in most cyanobacteria; the NdhL protein is part of a complex essential for cyanobacterial respiration. A cpld38 mutant of Chlamydomonas reinhardtii does not grow on minimal medium, is high light-sensitive under photoheterotrophic conditions, has lower accumulation of photosynthetic complexes, reduced photosynthetic electron flow to P700(+), and reduced photochemical efficiency of photosystem II (ΦPSII); these phenotypes are rescued by a wild-type copy of CPLD38. Single turnover flash experiments and biochemical analyses demonstrated that cytochrome b6f function was severely compromised, and the levels of transcripts and polypeptide subunits of the cytochrome b6f complex were also significantly lower in the cpld38 mutant. Furthermore, subunits of the cytochrome b6f complex in mutant cells turned over much more rapidly than in wild-type cells. Interestingly, PTOX2 and NDA2, two major proteins involved in chlororespiration, were more than 5-fold higher in mutants relative to wild-type cells, suggesting a shift in the cpld38 mutant from photosynthesis toward chlororespiratory metabolism, which is supported by experiments that quantify the reduction state of the plastoquinone pool. Together, these findings support the hypothesis that CPLD38 impacts the stability of the cytochrome b6f complex and possibly plays a role in balancing redox inputs to the quinone pool from photosynthesis and chlororespiration.
authors.affiliation: Department of Plant Biology, Carnegie Institute for Science, Stanford, California 94305, USA. mlh300@stanford.edu
authors.forename: Mark L
authors.initials: ML
authors.lastname: Heinnickel
authors.forename: Jean
authors.initials: J
authors.lastname: Alric
authors.forename: Tyler
authors.initials: T
authors.lastname: Wittkopp
authors.forename: Wenqiang
authors.initials: W
authors.lastname: Yang
authors.forename: Claudia
authors.initials: C
authors.lastname: Catalanotti
authors.forename: Rachel
authors.initials: R
authors.lastname: Dent
authors.forename: Krishna K
authors.initials: KK
authors.lastname: Niyogi
authors.forename: Francis-Andre
authors.initials: FA
authors.lastname: Wollman
authors.forename: Arthur R
authors.initials: AR
authors.lastname: Grossman
mesh_terms: D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism*; D002734:Chlorophyll / Q000378:metabolism; D025222:Chloroplast Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism; D045346:Cytochrome b6f Complex / Q000235:genetics / Q000378:metabolism*; D045347:Cytochromes b6 / Q000235:genetics / Q000378:metabolism; D045348:Cytochromes f / Q000235:genetics / Q000378:metabolism; D004579:Electron Transport; D015870:Gene Expression; D015151:Immunoblotting; D008027:Light; D009154:Mutation; D010084:Oxidation-Reduction; D010788:Photosynthesis / Q000235:genetics / Q000528:radiation effects; D045322:Photosynthetic Reaction Center Complex Proteins / Q000235:genetics / Q000378:metabolism*; D045331:Photosystem I Protein Complex / Q000235:genetics / Q000378:metabolism; D045332:Photosystem II Protein Complex / Q000235:genetics / Q000378:metabolism; D010971:Plastoquinone / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D060431:Thylakoid Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D020524:Thylakoids / Q000378:metabolism
chemical_list: D045347:Cytochromes b6; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D045332:Photosystem II Protein Complex; D060431:Thylakoid Membrane Proteins; D002734:Chlorophyll; D045346:Cytochrome b6f Complex; D045348:Cytochromes f; D025222:Chloroplast Proton-Translocating ATPases; D010971:Plastoquinone