A unique Rab GTPase, EhRabA, of Entamoeba histolytica, localizes to the leading edge of motile cells.
Entamoeba histolytica, an enteric protozoan parasite, infects 10% of the world's population leading to 50 million cases of invasive amoebiasis annually. Parasite vesicle trafficking and motility, which relies on vesicle trafficking to deliver membrane and membrane components to the leading edge, are important for virulence however little is known about the molecular mechanisms regulating these functions. Since Rab GTPases are known modulators of vesicle trafficking we have characterized a Rab GTPase of Entamoeba, EhRabA. Sequence analysis revealed that EhRabA shared limited homology with any known Rab suggesting that it is a novel member of this protein family. Immunofluorescence microscopy using EhRabA-specific antibodies demonstrated that EhRabA did not colocalize with markers for the Golgi apparatus, endoplasmic reticulum, pinosomes, or phagosomes. These data suggest that this Rab may not play a role in vesicle trafficking between these organelles. In quiescent Entamoeba cells, EhRabA localized to vesicles throughout the cytoplasm consistent with a role in vesicle trafficking, however, in motile cells this protein localized to small vesicles in the leading edge. In addition, when E. histolytica trophozoites were exposed to an N-formyl peptide (N-formylmethionylleucylphenylalanine) cell polarization, the formation of membrane extensions, and the translocation of EhRabA to these membrane extensions was observed. Taken together, these results suggest that EhRabA may function in the formation of membrane extensions perhaps by regulating the delivery of membrane and/or cell surface molecules to the plasma membrane.
Welter
Brenda H
BH
Department of Biological Sciences, 132 Long Hall, Clemson University, Clemson, SC 29634, USA.
Temesvari
Lesly A
LA
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D004748:Entamoeba histolytica / Q000201:enzymology* / Q000502:physiology; D008969:Molecular Sequence Data; D009068:Movement / Q000502:physiology; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D020691:rab GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D015800:Protozoan Proteins; D020691:rab GTP-Binding Proteins