Function and regulation of the Caenorhabditis elegans Rab32 family member GLO-1 in lysosome-related organelle biogenesis.

Caitlin Morris,Laura Voss,Youli Jian,Marie Harp,Simran Handa,Greg J Hermann,Chonglin Yang,Kimberly Peloza,My Van Vo,Olivia K Foster,Fiona M Rambo,Hannah Somhegyi,Kaveh Ashrafi

doi:10.1371/journal.pgen.1007772

Abstract

Cell type-specific modifications of conventional endosomal trafficking pathways lead to the formation of lysosome-related organelles (LROs). C. elegans gut granules are intestinally restricted LROs that coexist with conventional degradative lysosomes. The formation of gut granules requires the Rab32 family member GLO-1. We show that the loss of glo-1 leads to the mistrafficking of gut granule proteins but does not significantly alter conventional endolysosome biogenesis. GLO-3 directly binds to CCZ-1 and they both function to promote the gut granule association of GLO-1, strongly suggesting that together, GLO-3 and CCZ-1 activate GLO-1. We found that a point mutation in GLO-1 predicted to spontaneously activate, and function independently of it guanine nucleotide exchange factor (GEF), localizes to gut granules and partially restores gut granule protein localization in ccz-1(-) and glo-3(-) mutants. CCZ-1 forms a heterodimeric complex with SAND-1(MON1), which does not function in gut granule formation, to activate RAB-7 in trafficking pathways to conventional lysosomes. Therefore, our data suggest a model whereby the function of a Rab GEF can be altered by subunit exchange. glo-3(-) mutants, which retain low levels of GLO-3 activity, generate gut granules that lack GLO-1 and improperly accumulate RAB-7 in a SAND-1 dependent process. We show that GLO-1 and GLO-3 restrict the distribution of RAB-7 to conventional endolysosomes, providing insights into the segregation of pathways leading to conventional lysosomes and LROs.

Highlights

Caenorhabditis elegans gut granules are lysosome related organelles (LROs) [1], cell typerestricted compartments with diverse functions that share characteristics with conventional lysosomes [2]
We recently found that ccz-1(-) mutants lack gut granules, whereas rab-7(-) mutants have only a minor defect in gut granule protein localization, and sand-1 is dispensable for gut granule biogenesis [19]
To determine if GLO-3 can physically interact with CCZ-1, we screened for interactions between the two full-length proteins using the yeast two-hybrid system

Summary

Introduction

Caenorhabditis elegans gut granules are lysosome related organelles (LROs) [1], cell typerestricted compartments with diverse functions that share characteristics with conventional lysosomes [2]. These conspicuous intestine-specific compartments contain birefringent and autofluorescent material [3,4,5]. LRO biogenesis is mediated by evolutionarily conserved pathways that divert cargo away from conventional endosomes toward LROs [15, 16] Defects in these pathways cause Hermansky-Pudlak syndrome, a human condition characterized by a lack of dense granules and malformed melanosomes, LROs found within platelets and melanocytes, respectively [17]. Similar to many mammalian LROs, gut granule formation requires the BLOC-1 [18], HOPS [19], and AP-3 complexes [1], LYST [20], and the Rab family member GLO-1 [1]

Methods

Results

Conclusion