Expression of a gene for cyclophilin which contains an amino-terminal endoplasmic reticulum-targeting signal.
We isolated a novel gene for cyclophilin (CyP) first identified as an intracellular target of the immunosuppressant cyclosporin A and also known to have peptidyl-prolyl cis-trans isomerase (PPIase) activity, named ATCYP5 from Arabidopsis thaliana. ATCYP5 encoded a polypeptide with 201 amino acids with a putative ER-targeting signal sequence at its N-terminal, but without the typical ER-retention signal in its C-terminal. In addition, ATCYP5 protein contained a seven amino-acid long sequence which has been found previously only in cytosolic CyPs from plants. The synthetic mutant green fluorescent protein (sGFP; S65T) was fused to the N-terminal part of ATCYP5, and expressed in tobacco BY-2 cells. The fluorescence derived from the fusion protein was detected mainly around the nucleus, indicating translocation into ER. ATCYP5 was expressed mainly in young stems especially in the apical region and weakly in leaves and roots.
Saito
T
T
Research Institute of Molecular Genetics, Shimane University, Matsue, Japan.
Niwa
Y
Y
Ashida
H
H
Tanaka
K
K
Kawamukai
M
M
Matsuda
H
H
Nakagawa
T
T
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D002451:Cell Compartmentation; D004721:Endoplasmic Reticulum; D015870:Gene Expression; D015723:Gene Library; D017343:Genes, Plant; D017930:Genes, Reporter; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D019696:Peptidylprolyl Isomerase / Q000096:biosynthesis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D021382:Protein Sorting Signals / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D008164:Luminescent Proteins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005966:Glucuronidase; D019696:Peptidylprolyl Isomerase