Photosystem II assembly and repair are differentially localized in Chlamydomonas.
Many proteins of the photosynthesis complexes are encoded by the genome of the chloroplast and synthesized by bacterium-like ribosomes within this organelle. To determine where proteins are synthesized for the de novo assembly and repair of photosystem II (PSII) in the chloroplast of Chlamydomonas reinhardtii, we used fluorescence in situ hybridization, immunofluorescence staining, and confocal microscopy. These locations were defined as having colocalized chloroplast mRNAs encoding PSII subunits and proteins of the chloroplast translation machinery specifically under conditions of PSII subunit synthesis. The results revealed that the synthesis of the D1 subunit for the repair of photodamaged PSII complexes occurs in regions of the chloroplast with thylakoids, consistent with the current model. However, for de novo PSII assembly, PSII subunit synthesis was detected in discrete regions near the pyrenoid, termed T zones (for translation zones). In two PSII assembly mutants, unassembled D1 subunits and incompletely assembled PSII complexes localized around the pyrenoid, where we propose that they mark an intermediate compartment of PSII assembly. These results reveal a novel chloroplast compartment that houses de novo PSII biogenesis and the regulated transport of newly assembled PSII complexes to thylakoid membranes throughout the chloroplast.
Biology Department, Concordia University, Montreal, Quebec, Canada H4B 1R6.
James
J
Uniacke
William
W
Zerges
D020418:Algal Proteins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002696:Chlamydomonas / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000528:radiation effects; D008027:Light; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009154:Mutation / Q000235:genetics; D045332:Photosystem II Protein Complex / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000528:radiation effects; D021122:Protein Subunits / Q000096:biosynthesis; D021381:Protein Transport / Q000528:radiation effects; D034443:RNA Transport / Q000528:radiation effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D012269:Ribosomal Proteins / Q000378:metabolism; D012273:Ribulose-Bisphosphate Carboxylase / Q000378:metabolism; D020524:Thylakoids / Q000378:metabolism / Q000528:radiation effects
D020418:Algal Proteins; D045332:Photosystem II Protein Complex; D021122:Protein Subunits; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D012269:Ribosomal Proteins; C101625:RbcL protein, plastid; D012273:Ribulose-Bisphosphate Carboxylase