Cytoskeletal association is important for differential targeting of glucose transporter isoforms in Leishmania.
The major glucose transporter of the parasitic protozoan Leishmania enriettii exists in two isoforms, one of which (iso-1) localizes to the flagellar membrane, while the other (iso-2) localizes to the plasma membrane of the cell body, the pellicular membrane. These two isoforms differ only in their cytosolic NH2-terminal domains. Using immunoblots and immunofluorescence microscopy of detergent-extracted cytoskeletons, we have demonstrated that iso-2 associates with the microtubular cytoskeleton that underlies the cell body membrane, whereas the flagellar membrane isoform iso-1 does not associate with the cytoskeleton. Deletion mutants that remove the first 25 or more amino acids from iso-1 are retargeted from the flagellum to the pellicular membrane, suggesting that these deletions remove a signal required for flagellar targeting. Unlike the full-length iso-1 protein, these deletion mutants associate with the cytoskeleton. Our results suggest that cytoskeletal binding serves as an anchor to localize the iso-2 transporter within the pellicular membrane, and that the flagellar targeting signal of iso-1 diverts this transporter into the flagellar membrane and away from the pellicular microtubules.
Snapp
E L
EL
Department of Molecular Microbiology and Immunology, Oregon Health Sciences University, Portland, Oregon 97201, USA.
Landfear
S M
SM
D000818:Animals; D001665:Binding Sites; D002458:Cell Fractionation; D003599:Cytoskeleton / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology; D018985:Epitopes, B-Lymphocyte; D005407:Flagella / Q000378:metabolism; D051273:Glucose Transporter Type 2; D018313:Leishmania enriettii / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D009004:Monosaccharide Transport Proteins / Q000235:genetics / Q000378:metabolism*; D017830:Octoxynol / Q000494:pharmacology; D021382:Protein Sorting Signals / Q000378:metabolism; D051381:Rats; D012995:Solubility
D003902:Detergents; D018985:Epitopes, B-Lymphocyte; D051273:Glucose Transporter Type 2; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D021382:Protein Sorting Signals; D017830:Octoxynol