h-89 and phosphatidylinositol-4-phosphate

Vesicles formed by the COPI complex function in retrograde transport from the Golgi to the endoplasmic reticulum (ER). Phosphatidylinositol transfer protein beta (PITPbeta), an essential protein that possesses phosphatidylinositol (PtdIns) and phosphatidylcholine (PtdCho) lipid transfer activity is known to localise to the Golgi and ER but its role in these membrane systems is not clear. To examine the function of PITPbeta at the Golgi-ER interface, RNA interference (RNAi) was used to knockdown PITPbeta protein expression in HeLa cells. Depletion of PITPbeta leads to a decrease in PtdIns(4)P levels, compaction of the Golgi complex and protection from brefeldin-A-mediated dispersal to the ER. Using specific transport assays, we show that anterograde traffic is unaffected but that KDEL-receptor-dependent retrograde traffic is inhibited. This phenotype can be rescued by expression of wild-type PITPbeta but not by mutants defective in docking, PtdIns transfer and PtdCho transfer. These data demonstrate that the PtdIns and PtdCho exchange activity of PITPbeta is essential for COPI-mediated retrograde transport from the Golgi to the ER.

Topics: 1-Phosphatidylinositol 4-Kinase; Biological Transport; Brefeldin A; Coat Protein Complex I; Endoplasmic Reticulum; Gene Deletion; Gene Knockdown Techniques; Glycosphingolipids; Golgi Apparatus; HeLa Cells; Humans; Isoquinolines; Models, Biological; Mutant Proteins; Phosphatidylcholines; Phosphatidylinositol Phosphates; Phosphatidylinositols; Phospholipid Transfer Proteins; Protein Kinase Inhibitors; Receptors, Peptide; Sulfonamides