TBC-8, a putative RAB-2 GAP, regulates dense core vesicle maturation in Caenorhabditis elegans

PLoS Genet. 2012;8(5):e1002722. doi: 10.1371/journal.pgen.1002722. Epub 2012 May 24.

Abstract

Dense core vesicles (DCVs) are thought to be generated at the late Golgi apparatus as immature DCVs, which subsequently undergo a maturation process through clathrin-mediated membrane remodeling events. This maturation process is required for efficient processing of neuropeptides within DCVs and for removal of factors that would otherwise interfere with DCV release. Previously, we have shown that the GTPase, RAB-2, and its effector, RIC-19, are involved in DCV maturation in Caenorhabditis elegans motoneurons. In rab-2 mutants, specific cargo is lost from maturing DCVs and missorted into the endosomal/lysosomal degradation route. Cargo loss could be prevented by blocking endosomal delivery. This suggests that RAB-2 is involved in retention of DCV components during the sorting process at the Golgi-endosomal interface. To understand how RAB-2 activity is regulated at the Golgi, we screened for RAB-2-specific GTPase activating proteins (GAPs). We identified a potential RAB-2 GAP, TBC-8, which is exclusively expressed in neurons and which, when depleted, shows similar DCV maturation defects as rab-2 mutants. We could demonstrate that RAB-2 binds to its putative GAP, TBC-8. Interestingly, TBC-8 also binds to the RAB-2 effector, RIC-19. This interaction appears to be conserved as TBC-8 also interacted with the human ortholog of RIC-19, ICA69. Therefore, we propose that a dynamic ON/OFF cycling of RAB-2 at the Golgi induced by the GAP/effector complex is required for proper DCV maturation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autoantigens / metabolism
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / metabolism
  • Caenorhabditis elegans* / genetics
  • Caenorhabditis elegans* / growth & development
  • Clathrin-Coated Vesicles / genetics
  • Clathrin-Coated Vesicles / physiology
  • Endosomes / genetics
  • Endosomes / metabolism
  • GTPase-Activating Proteins / genetics
  • GTPase-Activating Proteins / metabolism*
  • Gene Expression Regulation
  • Golgi Apparatus* / genetics
  • Golgi Apparatus* / metabolism
  • Humans
  • Motor Neurons / metabolism
  • Protein Transport / genetics
  • Secretory Vesicles* / genetics
  • Secretory Vesicles* / physiology
  • rab2 GTP-Binding Protein / genetics*
  • rab2 GTP-Binding Protein / metabolism
  • rab3 GTP-Binding Proteins / genetics
  • rab3 GTP-Binding Proteins / metabolism

Substances

  • Autoantigens
  • Caenorhabditis elegans Proteins
  • GTPase-Activating Proteins
  • ric-19 protein, C elegans
  • TBC-8 protein, C elegans
  • rab2 GTP-Binding Protein
  • rab3 GTP-Binding Proteins