title: Genetic analysis of the cytoplasmic domain of the PfRh2b merozoite invasion protein of Plasmodium falciparum.
abstract: Apicomplexan parasites employ multiple adhesive ligands for recognition and entry into host cells. The Duffy binding-like (DBL) and the reticulocyte binding protein-like (RBL) families are central to the invasion of erythrocytes by the malaria parasite. These type-1 transmembrane proteins are composed of large ectodomains and small conserved cytoplasmic tail domains. The cytoplasmic tail domain of the micronemal DBL protein EBA-175 is required for a functional ligand-receptor interaction, but not for correct trafficking and localisation. Here we focus on the cytoplasmic tail domain of the rhoptry-localised Plasmodium falciparum RBL PfRh2b. We have identified a conserved sequence of six amino acids, enriched in acidic residues, in the cytoplasmic tail domains of RBL proteins from Plasmodium spp. Genetic analyses reveal that the entire cytoplasmic tail and the conserved motif within the cytoplasmic tail are indispensable for invasion P. falciparum. Site-directed mutagenesis of the conserved moiety reveals that changes in the order of the amino acids of the conserved moiety, but not the charge of the sequence, can be tolerated. Shuffling of the motif has no effect on either invasion phenotype or PfRh2b expression and trafficking. Although the PfRh2b gene can be readily disrupted, our results suggest that modification of the PfRh2b cytoplasmic tail results in strong dominant negative activity, highlighting important differences between the PfRh2b and EBA-175 invasion ligands.
authors.affiliation: Harvard School of Public Health, Boston, MA 02115, USA.
authors.forename: Tiffany M
authors.initials: TM
authors.lastname: DeSimone
authors.forename: Amy K
authors.initials: AK
authors.lastname: Bei
authors.forename: Cameron V
authors.initials: CV
authors.lastname: Jennings
authors.forename: Manoj T
authors.initials: MT
authors.lastname: Duraisingh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D003593:Cytoplasm / Q000737:chemistry* / Q000469:parasitology; D004912:Erythrocytes / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D052940:Merozoites / Q000469:parasitology; D008954:Models, Biological; D016297:Mutagenesis, Site-Directed; D010963:Plasmodium falciparum / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D015800:Protozoan Proteins / Q000235:genetics*
chemical_list: D002352:Carrier Proteins; D015800:Protozoan Proteins; C545352:Rh2b protein, Plasmodium falciparum