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.

Publication Date

6-10-2016

Publication Title

Journal of Biological Chemistry

Volume

291

Issue

24

Number of Pages

12514-12526

Document Type

Article

Personal Identifier

scopus

DOI Link

https://doi.org/10.1074/jbc.M115.705269

Socpus ID

84974528477 (Scopus)

Source API URL

https://api.elsevier.com/content/abstract/scopus_id/84974528477

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