The ArfGEF GBF-1 Is Required for ER Structure, Secretion and Endocytic Transport in C. elegans

PLoS One. 2013 Jun 19;8(6):e67076. doi: 10.1371/journal.pone.0067076. Print 2013.

Abstract

Small GTPases of the Sar/Arf family are essential to generate transport containers that mediate communication between organelles of the secretory pathway. Guanine nucleotide exchange factor (GEFs) activate the small GTPases and help their anchorage in the membrane. Thus, GEFs in a way temporally and spatially control Sar1/Arf1 GTPase activation. We investigated the role of the ArfGEF GBF-1 in C. elegans oocytes and intestinal epithelial cells. GBF-1 localizes to the cis-Golgi and is part of the t-ER-Golgi elements. GBF-1 is required for secretion and Golgi integrity. In addition, gbf-1(RNAi) causes the ER reticular structure to become dispersed, without destroying ER exit sites (ERES) because the ERES protein SEC-16 was still localized in distinct punctae at t-ER-Golgi units. Moreover, GBF-1 plays a role in receptor-mediated endocytosis in oocytes, without affecting recycling pathways. We find that both the yolk receptor RME-2 and the recycling endosome-associated RAB-11 localize similarly in control and gbf-1(RNAi) oocytes. While RAB5-positive early endosomes appear to be less prominent and the RAB-5 levels are reduced by gbf-1(RNAi) in the intestine, RAB-7-positive late endosomes were more abundant and formed aggregates and tubular structures. Our data suggest a role for GBF-1 in ER structure and endosomal traffic.

MeSH terms

  • Animals
  • Caenorhabditis elegans / genetics
  • Caenorhabditis elegans / metabolism*
  • Caenorhabditis elegans Proteins / genetics*
  • Caenorhabditis elegans Proteins / metabolism*
  • Cells, Cultured
  • Endocytosis
  • Endoplasmic Reticulum / metabolism
  • Endoplasmic Reticulum / physiology*
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism
  • Guanine Nucleotide Exchange Factors / genetics*
  • Guanine Nucleotide Exchange Factors / metabolism*
  • Intestinal Mucosa / metabolism
  • Intestines / cytology
  • Oocytes / metabolism
  • RNA Interference

Substances

  • Caenorhabditis elegans Proteins
  • GBF-1 protein, C elegans
  • Guanine Nucleotide Exchange Factors

Grants and funding

Funding provided by the Swiss National Science Foundation and the University of Basel. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.