title: TbFRP, a novel FYVE-domain containing phosphoinositide-binding Ras-like GTPase from trypanosomes.
abstract: Ras-like small GTPases are regulatory proteins that control multiple aspects of cellular function, and are particularly prevalent in vesicular transport. A proportion of GTPase paralogs appear restricted to certain eukaryote lineages, suggesting roles specific to a restricted lineage, and hence potentially reflecting adaptation to individual lifestyles or ecological niche. Here we describe the role of a GTPase, TbFRP, a FYVE domain N-terminally fused to a Ras-like GTPase, originally identified in Trypanosoma brucei. As FYVE-domains specifically bind phosphoinositol 3-phosphate (PI3P), which associates with endosomes, we suggest that TbFRP may unite phosphoinositide and small G protein endosomal signaling in trypanosomatids. TbFRP orthologs are present throughout the Euglenazoa suggesting that FRP has functions throughout the group. We show that the FYVE domain of TbFRP is functional in PI3P-dependent membrane targeting and localizes at the endosomal region. Further, while TbFRP is apparently non-essential, knockdown and immunochemical evidence indicates that TbFRP is rapidly cleaved upon synthesis, releasing the GTPase and FYVE-domains. Finally, TbFRP expression at both mRNA and protein levels is cell density-dependent. Together, these data suggest that TbFRP is an endocytic GTPase with a highly unusual mechanism of action that involves proteolysis of the nascent protein and membrane targeting via PI3P.
authors.affiliation: Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QP, UK.
authors.forename: Vincent O
authors.initials: VO
authors.lastname: Adung'a
authors.forename: Mark C
authors.initials: MC
authors.lastname: Field
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D056898:Euglenozoa / Q000145:classification / Q000201:enzymology / Q000235:genetics; D020558:GTP Phosphohydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018503:Genome, Protozoan / Q000235:genetics; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010716:Phosphatidylinositols / Q000378:metabolism*; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D059748:Proteolysis; D011817:Rabbits; D015398:Signal Transduction / Q000502:physiology; D014346:Trypanosoma brucei brucei / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D014351:Trypanosomatina / Q000145:classification / Q000201:enzymology / Q000235:genetics
chemical_list: D010716:Phosphatidylinositols; C483312:phosphoinositide 3-phosphate; D020558:GTP Phosphohydrolases