Characterization of a targeting motif for a flagellar membrane protein in Leishmania enriettii.
The surface membranes of eukaryotic flagella and cilia are contiguous with the plasma membrane. Despite the absence of obvious physical structures that could form a barrier between the two membrane domains, the lipid and protein compositions of flagella and cilia are distinct from the rest of the cell surface membrane. We have exploited a flagellar glucose transporter from the parasitic protozoan Leishmania enriettii as a model system to characterize the first targeting motif for a flagellar membrane protein in any eukaryotic organism. In this study, we demonstrate that the flagellar membrane-targeting motif is recognized by several species of Leishmania. Previously, we demonstrated that the 130 amino acid NH(2)-terminal cytoplasmic domain of isoform 1 glucose transporter was sufficient to target a nonflagellar integral membrane protein into the flagellar membrane. We have now determined that an essential flagellar targeting signal is located between amino acids 20 and 35 of the NH(2)-terminal domain. We have further analyzed the role of specific amino acids in this region by alanine replacement mutagenesis and determined that single amino acid substitutions did not abrogate targeting to the flagellar membrane. However, individual mutations located within a cluster of five contiguous amino acids, RTGTT, conferred differences in the degree of targeting to the flagellar membrane and the flagellar pocket, implying a role for these residues in the mechanism of flagellar trafficking.
Snapp
E L
EL
Department of Molecular Microbiology, Oregon Health Sciences University, Portland, Oregon 97201, USA.
Landfear
S M
SM
D000818:Animals; D005407:Flagella / Q000737:chemistry*; D005455:Fluorescent Antibody Technique; D018313:Leishmania enriettii / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics; D018613:Microscopy, Confocal; D009004:Monosaccharide Transport Proteins / Q000737:chemistry / Q000235:genetics; D016296:Mutagenesis; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D014162:Transfection
D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins