A chloroplast FKBP interacts with and affects the accumulation of Rieske subunit of cytochrome bf complex.
Immunophilins are intracellular receptors of the immunosuppressants cyclosporin A, FK506, and rapamycin. Although all immunophilins possess peptidyl-prolyl isomerase activity and are identified from a wide range of organisms, little is known about their cellular functions. We report the characterization and functional analysis of an FK506 and rapamycin-binding protein (AtFKBP13) from Arabidopsis. The AtFKBP13 protein is synthesized as a precursor that is imported into chloroplasts and processed to the mature form located in the thylakoid lumen, as shown by chloroplast import assays and Western blot analysis. Experiments show that AtFKBP13 is translocated across the thylakoid membrane by the DeltapH-dependent pathway. Yeast two-hybrid screening identified Rieske FeS protein, a subunit of the cytochrome bf complex in the photosynthetic electron transport chain, as an interacting partner for AtFKBP13. Both yeast two-hybrid and in vitro protein-protein interaction assays showed that the precursor, but not the mature form, of AtFKBP13 interacted with Rieske protein, suggesting that interaction between the two proteins occurs along the import pathway. When AtFKBP13 expression was suppressed by RNA interference method, the level of Rieske protein was significantly increased in the transgenic plants.
Gupta
Rajeev
R
Department of Plant and Microbial Biology, University of California, Berkeley 94720, USA.
Mould
Ruth M
RM
He
Zengyong
Z
Luan
Sheng
S
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D001665:Binding Sites; D002736:Chloroplasts / Q000378:metabolism*; D003573:Cytochrome b Group / Q000378:metabolism*; D045346:Cytochrome b6f Complex; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D004579:Electron Transport; D014450:Electron Transport Complex III; D006863:Hydrogen-Ion Concentration; D007506:Iron-Sulfur Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D011498:Protein Precursors / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D022021:Tacrolimus Binding Proteins / Q000378:metabolism*; D020524:Thylakoids / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D029681:Arabidopsis Proteins; D003573:Cytochrome b Group; D018076:DNA, Complementary; D018744:DNA, Plant; C517220:FKBP13 protein, Arabidopsis; D007506:Iron-Sulfur Proteins; D011498:Protein Precursors; C037989:Rieske iron-sulfur protein; C070133:FKBP-13; D045346:Cytochrome b6f Complex; D022021:Tacrolimus Binding Proteins; D014450:Electron Transport Complex III