Secretory and endocytic pathways converge in a dynamic endosomal system in a primitive protozoan.
Leishmania are a group of primitive eukaryotic trypanosomatid protozoa that are apically polarized with a flagellum at their anterior end. Surrounding the base of the flagellum is the flagellar reservoir that constitutes the site for endocytosis and exocytosis in these organisms. In the present study, we define a novel multivesicular tubular compartment involved in the intracellular trafficking of macromolecules in Leishmania. This dynamic structure appears to subtend the flagellar reservoir and extends towards the posterior end of the cell. Functional domains of several surface-expressed proteins, such as the gp63 glycosyl phosphatidyl inositol anchor and the 3'nucleotidase/nuclease transmembrane domain were fused to green fluorescent protein. These chimeric proteins were found to traffic through the secretory pathway and, while reaching their intended destinations, also accumulated within the intracellular tubular compartment. Using various compounds that are efficient fluid-phase markers used to track endocytosis in higher eukaryotes, we showed that this tubular compartment constitutes an important station in the endocytic pathway of these cells. Based on our functional observations of its role in the trafficking of expressed proteins and endocytosed markers, this compartment appears to have properties similar to endosomes of higher eukaryotes.
Cell Biology Section, Laboratory of Parasitic Diseases, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-0425, USA.
E
E
Ghedin
A
A
Debrabant
J C
JC
Engel
D M
DM
Dwyer
D000818:Animals; D002478:Cells, Cultured; D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000502:physiology* / Q000648:ultrastructure; D005407:Flagella / Q000502:physiology / Q000648:ultrastructure; D049452:Green Fluorescent Proteins; D007893:Leishmania donovani / Q000502:physiology* / Q000648:ultrastructure; D018320:Leishmania major / Q000502:physiology* / Q000648:ultrastructure; D008164:Luminescent Proteins / Q000378:metabolism; D015800:Protozoan Proteins / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014162:Transfection
D008164:Luminescent Proteins; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins