Identification and localization of rab6, separation of rab6 from ERD2 and implications for an 'unstacked' Golgi, in Plasmodium falciparum.
The rab6 gene product in mammalian cells and yeast is localized to and regulates protein transport in the medial and trans Golgi cisternae, as well as the trans Golgi network. We have identified a homologue in the malaria parasite Plasmodium falciparum which displays a rab-like sequence that is 62.4% identical to mammalian rab6. In addition the parasite gene (Pfrab6 gene) contains an N-terminal hydrophobic domain, unique to P. falciparum. Antibodies developed to Pfrab6 localize protein in 4-7 well-resolved sites in a ring-stage parasite, as detected by high resolution fluorescence microscopy. This suggests that there are multiple, distinct foci of medial/trans Golgi membranes in a ring. ERD2 is a cis Golgi marker in mammalian cells. The plasmodial homologue of ERD2 (PfERD2) is concentrated in a single perinuclear region in a ring-stage parasite. This site is distinct from the Pfrab6 membranes, indicating that early and late Golgi markers can be segregated in P. falciparum. Mammalian cells contain a single Golgi complex where cis medial and trans markers are tightly stacked in closely apposed cisternae. In P. falciparum-rings however, rab6-associated membranes are not invariably 'stacked' with an ERD2 structure. In immunoelectron microscopy studies, both the PfERD2- and Pfrab6-associated membranes appear tubulovesicular in nature, devoid of cisternal morphology. Hence the Golgi of ring stage parasites may comprise of multiple, 'unstacked' tubulovesicular clusters, suggesting a primitive organization of the organelle in Plasmodia.
Van Wye
J
J
Department of Microbiology and Immunology, Stanford University School of Medicine, California 94305, USA.
Ghori
N
N
Webster
P
P
Mitschler
R R
RR
Elmendorf
H G
HG
Haldar
K
K
D000595:Amino Acid Sequence; D000818:Animals; D015415:Biomarkers; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D003001:Cloning, Molecular; D019084:Fluorescent Antibody Technique, Indirect; D017125:Genes, Protozoan; D006056:Golgi Apparatus / Q000737:chemistry* / Q000648:ultrastructure; D008565:Membrane Proteins / Q000302:isolation & purification*; D008856:Microscopy, Fluorescence / Q000295:instrumentation; D016253:Microscopy, Immunoelectron; D008961:Models, Structural; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000737:chemistry* / Q000235:genetics / Q000254:growth & development / Q000648:ultrastructure; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D018000:Receptors, Peptide; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D020691:rab GTP-Binding Proteins; D018631:ras Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
D015415:Biomarkers; D002352:Carrier Proteins; C489305:ERD2 protein, Plasmodium; C064922:ERD2 protein, S cerevisiae; D008565:Membrane Proteins; D015800:Protozoan Proteins; C094135:Rab6 protein; D018000:Receptors, Peptide; D029701:Saccharomyces cerevisiae Proteins; D020691:rab GTP-Binding Proteins; D018631:ras Proteins