The membrane-bound histidine acid phosphatase TbMBAP1 is essential for endocytosis and membrane recycling in Trypanosoma brucei.
In the parasitic protozoan Trypanosoma brucei, endocytosis and exocytosis occur exclusively at an invagination of the plasma membrane around the base of the flagellum, called the flagellar pocket, which actively communicates by vesicular membrane flow with cisternal/tubulovesicular endosomes. The division of the cell surface into three morphologically distinct sub-domains and the rapid plasma membrane turnover establishes T. brucei as an interesting model for investigations on the sorting and recycling of membrane proteins. In this study we show that the type I membrane protein TbMBAP1, an L-(+)-tartrate-sensitive acid phosphatase, is present in all endosomal membranes but is virtually absent from the lysosome membrane (where this type of protein is mainly found in other organisms) and is not detectable at the cell surface. The endosomal localization of TbMBAP1 is a function of protein abundance. Moderate overexpression (three- to fourfold) leads to an increased appearance within the flagellar pocket membrane. At higher levels the protein is found in the flagellum, and routing to the pellicular plasma membrane is observed at levels 10- to 25-fold above that of wild type. In other organisms L-(+)-tartrate-sensitive acid phosphatases appear to be dispensable but TbMBAP1 is essential, as shown by RNA interference, which causes growth arrest followed by cell death. Comparison of the phenotype of TbMBAP1-depleted cells with that of cells in which endocytosis or exocytosis has been specifically inhibited by RNAi against clathrin of RAB11, reveals that TbMBAP1 is essential for both incoming and recycling membrane traffic. During differentiation of the organism from bloodstream to insect stage, TbMBAP1 is down-regulated and differentially modified in parallel with a 10-fold decrease in the rate of endocytosis.
Engstler
Markus
M
Ludwig-Maximilians-Universität, Department Biologie I, Genetik, Maria-Ward-Strasse 1a, München, 80638, Germany. engstler@lrz.uni-muenchen.de
Weise
Frank
F
Bopp
Karoline
K
Grünfelder
Christoph G
CG
Günzel
Mark
M
Heddergott
Niko
N
Overath
Peter
P
D000135:Acid Phosphatase / Q000378:metabolism*; D000818:Animals; D016923:Cell Death; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002966:Clathrin / Q000378:metabolism; D015536:Down-Regulation; D004705:Endocytosis; D011992:Endosomes / Q000201:enzymology; D005407:Flagella / Q000378:metabolism / Q000648:ultrastructure; D006639:Histidine / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D008958:Models, Molecular; D021381:Protein Transport; D000071681:Tartrate-Resistant Acid Phosphatase; D014346:Trypanosoma brucei brucei / Q000378:metabolism* / Q000648:ultrastructure; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D000099193:rab11 GTP-Binding Proteins
D000135:Acid Phosphatase; D002966:Clathrin; D006639:Histidine; D007527:Isoenzymes; D008565:Membrane Proteins; D000071681:Tartrate-Resistant Acid Phosphatase; D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins