Chloroplasts can accommodate inclusion bodies. Evidence from a mutant of Chlamydomonas reinhardtii defective in the assembly of the chloroplast ATP synthase.
We identified two neighboring missense mutations in the chloroplast atpA gene which are responsible for the defect of ATP synthase assembly in the FUD16 mutant from Chlamydomonas reinhardtii. The two corresponding amino acid substitutions, Ile184-->Asn and Asn186-->Tyr, occurred at strictly conserved sites among the alpha and beta subunits of (C)F1 complexes from bacteria, mitochondria, and chloroplasts. The altered region in the alpha polypeptide chain is located 7 amino acids downstream of the P-loop, which forms most of the conserved nucleotide binding site. Although the resulting chloroplast mutant fails to accumulate most of the ATP synthase subunits, it displays an increased intracellular content in both the alpha and beta subunits. We demonstrate that the two subunits do not bind to the thylakoid membranes but associate and overaccumulate in the chloroplast stroma as inclusion bodies. Increased rates of synthesis of the two subunits in the mutant point to an early interaction between the two subunits during their biogenesis.
Ketchner
S L
SL
Service de Photosynthèse, URA/CNRS 1187, Institut de Biologie Physico-Chimique, Paris, France.
Drapier
D
D
Olive
J
J
Gaudriault
S
S
Girard-Bascou
J
J
Wollman
F A
FA
D000595:Amino Acid Sequence; D000818:Animals; D001216:Asparagine; D001483:Base Sequence; D002499:Centrifugation, Density Gradient; D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure*; D002736:Chloroplasts / Q000201:enzymology* / Q000648:ultrastructure; D017124:Conserved Sequence; D003433:Crosses, Genetic; D005800:Genes, Fungal; D007150:Immunohistochemistry; D002479:Inclusion Bodies / Q000648:ultrastructure*; D007532:Isoleucine; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D010641:Phenotype; D017354:Point Mutation; D006180:Proton-Translocating ATPases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping; D014443:Tyrosine
D046911:Macromolecular Substances; D007532:Isoleucine; D014443:Tyrosine; D001216:Asparagine; D006180:Proton-Translocating ATPases