Novel variant of p230 trans-Golgi network protein identified by serum from Sjögren's syndrome patient.
Trans-Golgi network (TGN) protein p230 is a peripheral membrane protein associated with the cytoplasmic face of the TGN. TGNp230 is an extensively coiled-coil protein with flexible amino- and carboxyl-terminal ends, associates with non-clathrin-coated vesicles arising from the TGN, and is implicated in vesicle biogenesis. Here we used an autoimmune serum from a patient with S ogren's syndrome to clone partial cDNAs from a human hepatoma HepG2 expression library. The partial cDNAs encoded a novel amino-terminal splice variant of TGNp230. Specific reactivity of the autoimmune serum for p230 is supported by immunofluorescene staining of the Golgi apparatus, immunoblotting of a > 200-kDa HeLa cell protein, and reactivity with a bacterially expressed GST-p230 fusion protein. The alternative splicing occurs within the first proline-rich domain of p230. It comprises a deletion of 30 bp followed immediately by an additional 66 bp absent in the published sequence. RT-PCR analysis indicated that the splicing occurs independently of previously reported carboxyl-terminal splicing, and that this novel splice variant is more frequent than the previously reported p230. The novel splice variant of p230 is also located at the TGN. We propose that p230 splice variants may be implicated in selection of cargo molecules for vesicles arising from the TGN.
Tsukada
Y
Y
The Third Department of Internal Medicine, Gunma University School of Medicine, Japan.
Ichikawa
H
H
Chai
Z
Z
Lai
F P
FP
Dunster
K
K
Sentry
J W
JW
Toh
B H
BH
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D001323:Autoantibodies / Q000276:immunology*; D001324:Autoantigens / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D005260:Female; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012859:Sjogren's Syndrome / Q000097:blood / Q000276:immunology*; D013698:Templates, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
D001323:Autoantibodies; D001324:Autoantigens; C098772:GOLGA4 protein, human; D008565:Membrane Proteins; D012333:RNA, Messenger