title: Two Rab7 isotypes, EhRab7A and EhRab7B, play distinct roles in biogenesis of lysosomes and phagosomes in the enteric protozoan parasite Entamoeba histolytica.
abstract: Rab7 small GTPase plays a crucial role in the regulation of trafficking to late endosomes, lysosomes and phagosomes. While most eukaryotes encode a single Rab7, the parasitic protist Entamoeba histolytica possesses nine Rab7. In this study, to understand the significance of the presence of multiple Rab7 isotypes, a role of two representative Rab7 isotypes, EhRab7A and EhRab7B, was investigated. EhRab7B was exclusively localized to acidic vacuoles containing lysosomal proteins, e.g. amoebapore-A and cysteine protease. This lysosome localization of EhRab7B was in good contrast to EhRab7A, localized to a non-acidic compartment in steady state, and only partially colocalized with lysosomal proteins. Overexpression of EhRab7B resulted in augmentation of late endosome/lysosome acidification, similar to the EhRab7A overexpression. Expression of EhRab7B-GTP mutant caused dominant-negative phenotypes including decrease in late endosome/lysosome acidification and missecretion of lysosomal proteins, while EhRab7A-GTP enhanced acidification but did not affect either intracellular or secreted cysteine protease activity. Expression of either EhRab7B or EhRab7B-GTP mutant caused defect in phagocytosis, concomitant with the disturbed formation and disassembly of prephagosomal vacuoles, the compartment previously shown to be linked to efficient ingestion. Altogether, these data indicate that the two Rab7 isotypes play distinct but co-ordinated roles in lysosome and phagosome biogenesis.
authors.affiliation: Department of Parasitology, National Institute of Infectious Diseases, 1-23-1 Toyama, Tokyo 162-8640, Japan.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Saito-Nakano
authors.forename: Biswa Nath
authors.initials: BN
authors.lastname: Mitra
authors.forename: Kumiko
authors.initials: K
authors.lastname: Nakada-Tsukui
authors.forename: Dan
authors.initials: D
authors.lastname: Sato
authors.forename: Tomoyoshi
authors.initials: T
authors.lastname: Nozaki
mesh_terms: D000818:Animals; D004748:Entamoeba histolytica / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D020558:GTP Phosphohydrolases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism*; D010587:Phagocytosis; D010588:Phagosomes / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D000090802:rab7 GTP-Binding Proteins
chemical_list: D007527:Isoenzymes; D015800:Protozoan Proteins; D000090802:rab7 GTP-Binding Proteins; D020558:GTP Phosphohydrolases; C464925:Rab7 protein, Entamoeba histolytica