title: Silencing of Small Valosin-containing Protein-interacting Protein (SVIP) Reduces Very Low Density Lipoprotein (VLDL) Secretion from Rat Hepatocytes by Disrupting Its Endoplasmic Reticulum (ER)-to-Golgi Trafficking.
abstract: The transport of nascent very low density lipoprotein (VLDL) particles from the endoplasmic reticulum (ER) to the Golgi determines their secretion by the liver and is mediated by a specialized ER-derived vesicle, the VLDL transport vesicle (VTV). Our previous studies have shown that the formation of ER-derived VTV requires proteins in addition to coat complex II proteins. The VTV proteome revealed that a 9-kDa protein, small valosin-containing protein-interacting protein (SVIP), is uniquely present in these specialized vesicles. Our biochemical and morphological data indicate that the VTV contains SVIP. Using confocal microscopy and co-immunoprecipitation assays, we show that SVIP co-localizes with apolipoprotein B-100 (apoB100) and specifically interacts with VLDL apoB100 and coat complex II proteins. Treatment of ER membranes with myristic acid in the presence of cytosol increases SVIP recruitment to the ER in a concentration-dependent manner. Furthermore, we show that myristic acid treatment of hepatocytes increases both VTV budding and VLDL secretion. To determine the role of SVIP in VTV formation, we either blocked the SVIP protein using specific antibodies or silenced SVIP by siRNA in hepatocytes. Our results show that both blocking and silencing of SVIP lead to significant reduction in VTV formation. Additionally, we show that silencing of SVIP reduces VLDL secretion, suggesting a physiological role of SVIP in intracellular VLDL trafficking and secretion. We conclude that SVIP acts as a novel regulator of VTV formation by interacting with its cargo and coat proteins and has significant implications in VLDL secretion by hepatocytes.
authors.affiliation: Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, Florida 32827.
authors.forename: Samata
authors.initials: S
authors.lastname: Tiwari
authors.affiliation: Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, Florida 32827.
authors.forename: Shaila
authors.initials: S
authors.lastname: Siddiqi
authors.affiliation: Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, Florida 32827.
authors.forename: Olga
authors.initials: O
authors.lastname: Zhelyabovska
authors.affiliation: Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, Florida 32827. Electronic address: shadab.siddiqi@ucf.edu.
authors.forename: Shadab A
authors.initials: SA
authors.lastname: Siddiqi
mesh_terms: D000818:Animals; D053299:Apolipoprotein B-100 / Q000378:metabolism; D002478:Cells, Cultured; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015151:Immunoblotting; D008079:Lipoproteins, VLDL / Q000378:metabolism*; D008297:Male; D018613:Microscopy, Confocal; D019814:Myristic Acid / Q000494:pharmacology; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D034622:RNA Interference; D017207:Rats, Sprague-Dawley; D022161:Transport Vesicles / Q000378:metabolism; D014280:Triglycerides / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000378:metabolism
chemical_list: D053299:Apolipoprotein B-100; D008079:Lipoproteins, VLDL; D009687:Nuclear Proteins; C475849:SVIP protein, rat; D014280:Triglycerides; D033921:Vesicular Transport Proteins; D019814:Myristic Acid
keywords: COPII; VLDL transport vesicle (VTV); endoplasmic reticulum (ER); lipoprotein secretion; small VCP-interacting protein (SVIP); triglyceride; valosin-containing protein (VCP); very low density lipoprotein (VLDL); vesicles