Identification of the Plasmodium falciparum rhoptry neck protein 5 (PfRON5)

Curtidor, Hernando; Patiño, Liliana Catherine; Arévalo-Pinzón, Gabriela; Patarroyo, Manuel Elkín; Patarroyo, Manuel A.

doi:10.1016/j.gene.2010.12.005

Cited by 22 publications

(30 citation statements)

References 57 publications

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“…SignalP 4.1 predicted a putative endoplasmic reticulum (ER) transport signal peptide at aa 1−20 at the PyRON5 N-terminus (Fig. 1) similar to TgRON5, PfRON5 [5,32] and TeRON5, suggesting that RON5 proteins are targeted to the secretory pathway. TM domains were not predicted by Phobius and OCTOPUS for PyRON5.…”

Section: Determination Of Pyron5 Gene and Pyron5 Protein Primary Strumentioning

confidence: 99%

Expression and localization of rhoptry neck protein 5 in merozoites and sporozoites of Plasmodium yoelii

Mutungi

Yahata

Sakaguchi

et al. 2014

Parasitology International

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Host cell invasion by Apicomplexan parasites marks a crucial step in disease establishment and pathogenesis. The moving junction (MJ) is a conserved and essential feature among parasites of this phylum during host cell invasion, thus proteins that associate at this MJ are potential targets of drug and vaccine development. In both Toxoplasma gondii and Plasmodium falciparum, a micronemal protein, Apical Membrane Antigen 1 (AMA1), and Rhoptry Neck proteins (RONs; RON2 and RON4) form an essential complex at the MJ. A new RON member, RON5, was shown to be important to stabilize RON2 during development and to associate with the MJ complex in T. gondii and also to be immunoprecipitated by anti-AMA1 antibody in P. falciparum. However, the detailed molecular nature of RON5 in Plasmodium is not well understood. In this study, Plasmodium yoelii RON5 gene (pyron5) was identified as an ortholog of P. falciparum and Plasmodium berghei ron5. The pyron5 exon-intron structure was validated by comparing genomic DNA sequences and experimentally determining fulllength complementary DNA sequence. PyRON5 was detected in water-insoluble fractions but no reliable transmembrane domain(s) were predicted by transmembrane prediction algorithms. PyRON5 formed a complex with PyRON4, PyRON2, and PyAMA1 in late schizont protein extract. Taken together, we infer that these results suggest that PyRON5 associates with membrane indirectly via other MJ components. Indirect immunofluorescence assay and immunoelectron microscopy localized PyRON5 at the rhoptry neck of the late schizont merozoites and at the rhoptry of sporozoites. The two-stage expression of PyRON5 suggests that PyRON5 plays roles in invasion of not only erythrocytes, but also of mosquito salivary glands and/or mammalian hepatocytes. Graphic Abstract Highlights• PyRON5 forms a complex with RON2, RON4, and AMA1 in schizont extract.• PyRON5, a putative soluble protein, exists in the water-insoluble fractions.• These suggest that PyRON5 associates with membrane via other complex component(s).• PyRON5 is expressed in the rhoptry in both merozoite and sporozoite parasites.• Targeting PyRON5 may prevent cell invasion by both merozoite and sporozoite. Mutungi et al., 2

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Section: Determination Of Pyron5 Gene and Pyron5 Protein Primary Strumentioning

confidence: 99%

Expression and localization of rhoptry neck protein 5 in merozoites and sporozoites of Plasmodium yoelii

Mutungi

Yahata

Sakaguchi

et al. 2014

Parasitology International

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“…Pf RON2 [21], Pf RON4 [22] and Pf RON5 [23] are located in the rhoptry neck and co-immunoprecipitate with Pf AMA-1 [21,22,24,25]. Furthermore, the Pf RON2 protein and the Pf AMA-1 ectodomain interaction has already been characterized, as well as its importance for erythrocyte invasion, suggesting that the mechanism described in T. gondii could be conserved among different members of the phylum Apicomplexa [19].…”

Section: Introductionmentioning

confidence: 99%

Pv RON2, a new Plasmodium vivax rhoptry neck antigen

Arévalo-Pinzón

Curtidor

Patiño

et al. 2011

Malar J

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BackgroundRhoptries are specialized organelles from parasites belonging to the phylum Apicomplexa; they secrete their protein content during invasion of host target cells and are sorted into discrete subcompartments within rhoptry neck or bulb. This distribution is associated with these proteins' role in tight junction (TJ) and parasitophorous vacuole (PV) formation, respectively.MethodsPlasmodium falciparum RON2 amino acid sequence was used as bait for screening the codifying gene for the homologous protein in the Plasmodium vivax genome. Gene synteny, as well as identity and similarity values, were determined for ron2 and its flanking genes among P. falciparum, P. vivax and other malarial parasite genomes available at PlasmoDB and Sanger Institute databases. Pvron2 gene transcription was determined by RT-PCR of cDNA obtained from the P. vivax VCG-1 strain. Protein expression and localization were assessed by Western blot and immunofluorescence using polyclonal anti-PvRON2 antibodies. Co-localization was confirmed using antibodies directed towards specific microneme and rhoptry neck proteins.Results and discussionThe first P. vivax rhoptry neck protein (named here PvRON2) has been identified in this study. PvRON2 is a 2,204 residue-long protein encoded by a single 6,615 bp exon containing a hydrophobic signal sequence towards the amino-terminus, a transmembrane domain towards the carboxy-terminus and two coiled coil α-helical motifs; these are characteristic features of several previously described vaccine candidates against malaria. This protein also contains two tandem repeats within the interspecies variable sequence possibly involved in evading a host's immune system. PvRON2 is expressed in late schizonts and localized in rhoptry necks similar to what has been reported for PfRON2, which suggests its participation during target cell invasion.ConclusionsThe identification and partial characterization of the first P. vivax rhoptry neck protein are described in the present study. This protein is homologous to PfRON2 which has previously been shown to be associated with PfAMA-1, suggesting a similar role for PvRON2.

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“…A previously described ELISA test was used for determining inoculated peptide immunogenicity [19]. Briefly, each of the inoculated peptides was sown in 96-well ELISA plates and incubated at 1:100 dilution of each serum.…”

Section: Methodsmentioning

confidence: 99%

“…Proteomics analysis of T. gondii rhoptries has shown minimum overlapping with P. falciparum proteins, except for some proteins localized in the rhoptry neck (RON), which could be involved in functions common to both parasites [16]. RON2 [17], RON4 [18] and RON5 [19] proteins have been identified to date in P. falciparum , forming part of a high molecular weight complex interacting with a microneme-derived protein called apical membrane antigen 1 (AMA-1) [20]. The assembly of this macromolecular complex allows TJ formation or irreversible binding which is important since this acts as a foothold for the parasite’s motor machinery to propel the parasite into the PV for successful invasion.…”

Section: Introductionmentioning

confidence: 99%

Annotation and characterization of the Plasmodium vivax rhoptry neck protein 4 (Pv RON4)

Arévalo-Pinzón

Curtidor

Abril

et al. 2013

Malar J

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BackgroundThe tight junction (TJ) is one of the most important structures established during merozoite invasion of host cells and a large amount of proteins stored in Toxoplasma and Plasmodium parasites’ apical organelles are involved in forming the TJ. Plasmodium falciparum and Toxoplasma gondii apical membrane antigen 1 (AMA-1) and rhoptry neck proteins (RONs) are the two main TJ components. It has been shown that RON4 plays an essential role during merozoite and sporozoite invasion to target cells. This study has focused on characterizing a novel Plasmodium vivax rhoptry protein, RON4, which is homologous to PfRON4 and PkRON4.MethodsThe ron4 gene was re-annotated in the P. vivax genome using various bioinformatics tools and taking PfRON4 and PkRON4 amino acid sequences as templates. Gene synteny, as well as identity and similarity values between open reading frames (ORFs) belonging to the three species were assessed. The gene transcription of pvron4, and the expression and localization of the encoded protein were also determined in the VCG-1 strain by molecular and immunological studies. Nucleotide and amino acid sequences obtained for pvron4 in VCG-1 were compared to those from strains coming from different geographical areas.ResultsPvRON4 is a 733 amino acid long protein, which is encoded by three exons, having similar transcription and translation patterns to those reported for its homologue, PfRON4. Sequencing PvRON4 from the VCG-1 strain and comparing it to P. vivax strains from different geographical locations has shown two conserved regions separated by a low complexity variable region, possibly acting as a “smokescreen”. PvRON4 contains a predicted signal sequence, a coiled-coil α-helical motif, two tandem repeats and six conserved cysteines towards the carboxy-terminus and is a soluble protein lacking predicted transmembranal domains or a GPI anchor. Indirect immunofluorescence assays have shown that PvRON4 is expressed at the apical end of schizonts and co-localizes at the rhoptry neck with PvRON2.ConclusionsGenomic, transcriptional and expression data reported for PvRON4, as well as its primary structure characteristics suggest that this protein participates in reticulocyte invasion, as has been shown for its homologue PfRON4.

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Identification of the Plasmodium falciparum rhoptry neck protein 5 (PfRON5)

Cited by 22 publications

References 57 publications

Expression and localization of rhoptry neck protein 5 in merozoites and sporozoites of Plasmodium yoelii

Expression and localization of rhoptry neck protein 5 in merozoites and sporozoites of Plasmodium yoelii

Pv RON2, a new Plasmodium vivax rhoptry neck antigen

Annotation and characterization of the Plasmodium vivax rhoptry neck protein 4 (Pv RON4)

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