The Secretory Protein Repertoire and Expanded Gene Families of Toxoplasma gondii and Other Apicomplexa

2007 ◽  
pp. 445-463
Author(s):  
T.J. Templeton
2015 ◽  
Vol 10 (2) ◽  
Author(s):  
Wayan T. Artama ◽  
Yulia Sari ◽  
Didik Tulus Subekti ◽  
Soenarwan Hery Poerwanto ◽  
Jarot Subandono

Rhoptry protein belongs to an excretory and secretory antigens (ESAs) that play an important role during activepenetration of parasite into the cell target. This protein an able Toxoplasma gondii to actively penetrate targetedcell, meanwhile ESAs protein stimulates intracellular vacuole modification. It is, therefore, after the parasitesuccessfully enter the cell target then Granule (GRA) proteins are responsible for the formation of parasitophorusvacuole, which is protect the fusion with other intracellular compartments such as lysosomal vacuole. Consequently,this parasite is being able to survive and multiply at the cell target. The current study was aimed to clone andsequens cDNA encoding for ROP-2 of local isolated T. gondii tachizoite through DNA recombinant technique.Total ribonucleic acid (RNA) was isolated from tachyzoites of local isolated T. gondii that were grown up in Balb/c mice. Messenger RNA was isolated from total RNA using PolyAtract mRNA Isolation System. Messenger RNA wasused as a template for synthesis cDNA using Riboclone cDNA Synthesis System AMV-RT. EcoRI adaptor fromRiboclone EcoRI Adaptor Ligation System was added to Complementary DNA and than ligated to pUC19. Recombinantplasmid was transformed into E. coli (XL1-Blue). The transformed E. coli XL-1 Blue were plated on LB agarcontaining X-Gal, IPTG and ampicillin. Recombinant clones (white colony) were picked up and grown up in theLB medium at 37oC overnight. Expression of recombinant protein was analysed by immunoblotting in order toidentify cDNA recombinant wich is express ESA of T. gondii local isolate. Recombinant plasmid were isolatedusing alkalilysis method and were elektroforated in 1% agarose gel. The isolated DNA recombinant plasmid wascut using Eco RI and then sequenced through Big Dye Terminator Mix AB1 377A Sequencer using M13 Forward andM13 Reverse primers. The conclusion of this results showed that the recombinant clone was coding for excretoryand secretory protein which has molecular weight of 54 kDa. The DNA alignments of sequence from the clonedgene showed 97% homology with gene encoding for ROP-2 of T. gondii RH isolate.Keywords: Toxoplasma gondii, tachizoite, ESA, complementary DNA, ROP2


2021 ◽  
Vol 35 (2) ◽  
Author(s):  
Shuang Li ◽  
Jing Liu ◽  
Heng Zhang ◽  
Zhepeng Sun ◽  
Zhu Ying ◽  
...  

2011 ◽  
Vol 80 (2) ◽  
pp. 476-482 ◽  
Author(s):  
Eric Y. Denkers ◽  
David J. Bzik ◽  
Barbara A. Fox ◽  
Barbara A. Butcher

ABSTRACTThe intracellular protozoanToxoplasma gondiiis well known for its skill at invading and living within host cells. New discoveries are now also revealing the astounding ability of the parasite to inject effector proteins into the cytoplasm to seize control of the host cell. This review summarizes recent advances in our understanding of one such secretory protein called ROP16. This molecule is released from rhoptries into the host cell during invasion. The ROP16 molecule acts as a kinase, directly activating both signal transducer and activator of transcription 3 (STAT3) and STAT6 signaling pathways. In macrophages, an important and preferential target cell of parasite infection, the injection of ROP16 has multiple consequences, including downregulation of proinflammatory cytokine signaling and macrophage deviation to an alternatively activated phenotype.


mBio ◽  
2012 ◽  
Vol 3 (6) ◽  
Author(s):  
James D. Wasmuth ◽  
Viviana Pszenny ◽  
Simon Haile ◽  
Emily M. Jansen ◽  
Alexandra T. Gast ◽  
...  

ABSTRACTTheToxoplasma gondiiSRSgene superfamily is structurally related toSRS29B(formerlySAG1), a surface adhesin that binds host cells and stimulates host immunity. Comparative genomic analyses of threeToxoplasmastrains identified 182SRSgenes distributed across 14 chromosomes at 57 genomic loci. Eight distinctSRSsubfamilies were resolved. A core 69 functional gene orthologs were identified, and strain-specific expansions and pseudogenization were common. Gene expression profiling demonstrated differential expression ofSRSgenes in a developmental-stage- and strain-specific fashion and identified nineSRSgenes as priority targets for gene deletion among the tissue-encysting coccidia. A Δsag1 ∆sag2Amutant was significantly attenuated in murine acute virulence and showed upregulated SRS29C (formerly SRS2) expression. Transgenic overexpression of SRS29C in the virulent RH parent was similarly attenuated. Together, these findings reveal SRS29C to be an important regulator of acute virulence in mice and demonstrate the power of integrated genomic analysis to guide experimental investigations.IMPORTANCEParasitic species employ large gene families to subvert host immunity to enable pathogen colonization and cause disease.Toxoplasma gondiicontains a large surface coat gene superfamily that encodes adhesins and virulence factors that facilitate infection in susceptible hosts. We generated an integrated bioinformatic resource to predict which genes from within this 182-gene superfamily of adhesin-encoding genes play an essential role in the host-pathogen interaction. Targeted gene deletion experiments with predicted candidate surface antigens identified SRS29C as an important negative regulator of acute virulence in murine models ofToxoplasmainfection. Our integrated computational and experimental approach provides a comprehensive framework, or road map, for the assembly and discovery of additional key pathogenesis genes contained within other large surface coat gene superfamilies from a broad array of eukaryotic pathogens.


2012 ◽  
Vol 12 (1) ◽  
pp. 30 ◽  
Author(s):  
Yap Boon Wooi Tommy ◽  
Theam Soon Lim ◽  
Rahmah Noordin ◽  
Geita Saadatnia ◽  
Yee Siew Choong

BMC Genomics ◽  
2015 ◽  
Vol 16 (1) ◽  
Author(s):  
Adrian B Hehl ◽  
Walter U Basso ◽  
Christoph Lippuner ◽  
Chandra Ramakrishnan ◽  
Michal Okoniewski ◽  
...  

1999 ◽  
Vol 112 (16) ◽  
pp. 2631-2638 ◽  
Author(s):  
K.M. Hager ◽  
B. Striepen ◽  
L.G. Tilney ◽  
D.S. Roos

Morphological examination of the highly polarized protozoan parasite Toxoplasma gondii suggests that secretory traffic in this organism progresses from the endoplasmic reticulum to the Golgi apparatus using the nuclear envelope as an intermediate compartment. While the endoplasmic reticulum is predominantly located near the basal end of the parasite, the Golgi is invariably adjacent to the apical end of the nucleus, and the space between the Golgi and nuclear envelope is filled with numerous coatomer-coated vesicles. Staining with antiserum raised against recombinant T. gondii beta-COP confirms its association with the apical juxtanuclear region. Perturbation of protein secretion using brefeldin A, microtubule inhibitors or dithiothreitol disrupts the Golgi, causing swelling of the nuclear envelope, particularly at its basal end. Prolonged drug treatment leads to gross distention of the endoplasmic reticulum, filling the basal end of the parasite. Cloning and sequencing of the T. gondii homolog of the chaperonin protein BiP identifies the carboxy-terminal amino acid sequence HDEL as this organism's endoplasmic reticulum-retention signal. Appending the HDEL motif to a recombinant secretory protein (a chimera between the parasite's major surface protein fusion, P30, and the Green Fluorescent Protein) causes this secretory reporter to be retained intracellularly. P30-GFP-HDEL fluorescence was most intense within the nuclear envelope, particularly at the apical end. These data support a model of secretion in which protein traffic from the endoplasmic reticulum to Golgi occurs via the apical end of the nuclear envelope.


2021 ◽  
Author(s):  
Terence Charles Theisen ◽  
John C. Boothroyd

Toxoplasma gondii has numerous, large, paralogous gene families that are likely critical for supporting its unparalleled host range: nearly any nucleated cell in almost any warm-blooded animal. The SRS (SAG1-related sequence) gene family encodes over 100 proteins, the most abundant of which are thought to be involved in parasite attachment and, based on their stage-specific expression, evading the host immune response. For most SRS proteins, however, little is understood about their function and expression profile. Single-parasite RNA-sequencing previously demonstrated that across an entire population of lab-grown tachyzoites, transcripts for over 70 SRS genes were detected in at least one parasite. In any one parasite, however, transcripts for an average of only 7 SRS genes were detected, two of which, SAG1 and SAG2A , were extremely abundant and detected in virtually all. These data do not address whether this pattern of sporadic SRS gene expression is consistently inherited among the progeny of a given parasite or arises independently of lineage. We hypothesized that if SRS expression signatures are stably inherited by progeny, subclones isolated from a cloned parent would be more alike in their expression signatures than they are to the offspring of another clone. In this report, we compare transcriptomes of clonally derived parasites to determine the degree to which expression of the SRS family is stably inherited in individual parasites. Our data indicate that in RH tachyzoites, SRS genes are variably expressed even between parasite samples subcloned from the same parent within approximately 10 parasite divisions (72 hours). This suggests that the pattern of sporadically expressed SRS genes is highly variable and not driven by inheritance mechanisms, at least under our conditions.


2012 ◽  
Vol 154 (6) ◽  
pp. 251-255 ◽  
Author(s):  
F. C. Frey ◽  
E. A. Berger-Schoch ◽  
C. D. Herrmann ◽  
G. Schares ◽  
N. Müller ◽  
...  
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