PIGEA-14, a novel coiled-coil protein affecting the intracellular distribution of polycystin-2.
Employing a yeast two-hybrid screen with the COOH terminus of polycystin-2, one of the proteins mutated in patients with polycystic kidney disease, we were able to isolate a novel protein that we call PIGEA-14 (polycystin-2 interactor, Golgi- and endoplasmic reticulum-associated protein with a molecular mass of 14 kDa). Molecular modeling only predicts a coiled-coil motif, but no other functional domains, in PIGEA-14. In a subsequent two-hybrid screen using PIGEA-14 as a bait, we found GM130, a component of the cis-compartment of the Golgi apparatus. Co-expression of the PIGEA-14 and PKD2 cDNAs in LLC-PK(1) and HeLa cells resulted in a redistribution of PIGEA-14 and polycystin-2 to the trans-Golgi network, which suggests that PIGEA-14 plays an important role in regulating the intracellular location of polycystin-2 and possibly other intracellular proteins. Our results also indicate that the intracellular trafficking of polycystin-2 is regulated both at the level of the endo-plasmic reticulum and that of the trans-Golgi network.
Hidaka
Sumi
S
Institute for Molecular and Cellular Anatomy, University of Regensburg, Universitätsstrasse 31, 93053 Regensburg, Germany.
Könecke
Vera
V
Osten
Larissa
L
Witzgall
Ralph
R
D000199:Actins / Q000737:chemistry; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D017353:Gene Deletion; D005982:Glutathione Transferase / Q000378:metabolism; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D018374:LLC-PK1 Cells; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D007690:Polycystic Kidney Diseases / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D013347:Subcellular Fractions / Q000378:metabolism; D050396:TRPP Cation Channels; D014162:Transfection; D020798:Two-Hybrid System Techniques
D000199:Actins; D002352:Carrier Proteins; D018076:DNA, Complementary; D005982:Glutathione Transferase; D008565:Membrane Proteins; D009687:Nuclear Proteins; D050396:TRPP Cation Channels; C473750:CBY1 protein, human; C490378:Cby1 protein, mouse