Trypanosoma cruzi genome project: biological characteristics and molecular typing of clone CL Brener

Zingales, Bianca; Pereira, M. E. S.; Oliveira, Riva P.; Almeida, Katia A.; Umezawa, Eufrosina Setsu; Souto, Ricardo Peres do; Vargas, Nancy; Cano, Maria Isabel Nogueira; Silveira, José Franco da; Nehme, N. S.; Morel, Carlos Médicis; Brener, Z.; Macedo, Andréa Mara

doi:10.1016/s0001-706x(97)00088-0

Cited by 75 publications

(47 citation statements)

References 25 publications

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“…Using nonphagocytic cells, we observed that trypomastigotes interact with autophagosomes at early times after infection and that autophagy and this strain presents important characteristics such as its ability to infect mice, its preferential parasitism of heart and muscle cells, and its high susceptibility to drugs used clinically in Chagas disease. 33 Although in this paper we mainly worked with the CL Brener clone, we also analyzed other clinically important T. cruzi strains such as T. cruzi RA and K98 which were initially isolated from patients with an acute and chronic Chagas disease, respectively. 34,35 Our results indicate that the parasitophorous vacuoles containing these strains also associate with the GFP-LC3 protein, indicating that the interaction of this parasite with the autophagic pathway is likely a general phenomenon.…”

Section: Resultsmentioning

confidence: 99%

“…We next studied the effect of the host-cell autophagy during the infection course of T. cruzi. According to the intracellular cycle time-span of this parasite, 33 we performed three experimental approaches to analyze the invasion, the differentiation and the replication stage, in cells subjected to conditions that stimulate autophagy such as amino acid deprivation or rapamycin. To test the invasion process CHO GFP-LC3 and CHO GFP (Vector) cells were preincubated in full nutrient or starvation media for 2 h and then infected with TCT (MOI = 50) in the same conditions, for 1 h before fixation (see diagram in Fig.…”

Section: Resultsmentioning

confidence: 99%

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The autophagic pathway is a key component in the lysosomal dependent entry ofTrypanosoma cruziinto the host cell

Romano

Arboit²,

Vázquez³

et al. 2009

Autophagy

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

The autophagic pathway is a key component in the lysosomal dependent entry ofTrypanosoma cruziinto the host cell

Romano

Arboit²,

Vázquez³

et al. 2009

Autophagy

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“…Culture conditions for T. cruzi CL-Brener cloned stock (38), parasite transfections parameters, and hybridization experiments were according to Di Noia et al (19), with modifications described in SI Text. Oligonucleotide sequences used in this work are listed in SI Table 3.…”

Section: Methodsmentioning

confidence: 99%

mRNA maturation by two-step trans-splicing/polyadenylation processing in trypanosomes

Jäger

Gaudenzi

Cassola

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Trypanosomes are unique eukaryotic cells, in that they virtually lack mechanisms to control gene expression at the transcriptional level. These microorganisms mostly control protein synthesis by posttranscriptional regulation processes, like mRNA stabilization and degradation. Transcription in these cells is polycistronic. Tens to hundreds of protein-coding genes of unrelated function are arrayed in long clusters on the same DNA strand. Polycistrons are cotranscriptionally processed by trans-splicing at the 5 end and polyadenylation at the 3 end, generating monocistronic units ready for degradation or translation. In this work, we show that some trans-splicing/polyadenylation sites may be skipped during normal polycistronic processing. As a consequence, dicistronic units or monocistronic transcripts having long 3 UTRs are produced. Interestingly, these unspliced transcripts can be processed into mature mRNAs by the conventional trans-splicing/ polyadenylation events leading to translation. To our knowledge, this is a previously undescribed mRNA maturation by trans-splicing uncoupled from transcription. We identified an RNA-recognition motif-type protein, homologous to the mammalian polypyrimidine tract-binding protein, interacting with one of the partially processed RNAs analyzed here that might be involved in exon skipping. We propose that splice-site skipping might be part of a posttranscriptional mechanism to regulate gene expression in trypanosomes, through the generation of premature nontranslatable RNA molecules.gene expression ͉ posttranscriptional regulation ͉ RNA processing R egulation of gene expression is central in defining the phenotype of a cell or organism. In the vast majority of cases, this regulation is controlled by transcriptional regulation in which DNA sequence elements, together with DNA-binding proteins, target genes for transcription by RNA polymerase II. In addition to regulating single genes, master regulators of transcription may give rise to a gene expression phenotype, a trait that may be inherited (1). Mature transcripts can also be regulated in their expression at the posttranscriptional level. A number of cis-motifs mainly located in the 5Ј and 3Ј UTRs determine the fate of the transcript by the use of highly specific and sometimes evolutionarily conserved machineries. This process is essentially achieved through RNA-binding proteins (RBPs) forming, together with the mRNA, ribonucleoprotein (RNP) complexes. It is the composition of the RNP complex that determines whether an mRNA is transported to the cytoplasm for translation, degradation, or other processing events (2, 3).Posttranscriptional regulation is common among Kinetoplastid parasites, like trypanosomes and Leishmania (4, 5), which are the causative agents of several infections affecting both humans and domestic animals worldwide (6). In these parasites, transcription by RNA polymerase II starts at a few genomic locations within chromosomes. In contrast to operons in bacteria, polycistronic units in trypanosomatids requir...

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“…Parasites-The CL Brener genome project reference clone and the Y strain were used throughout this study (33). Distinct T. cruzi stages (CL Brener clone) were obtained as described (20).…”

Section: Methodsmentioning

confidence: 99%

The Surface Coat of the Mammal-dwelling Infective Trypomastigote Stage of Trypanosoma cruzi Is Formed by Highly Diverse Immunogenic Mucins

Buscaglia

Campo

Noia

et al. 2004

Journal of Biological Chemistry

115

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A thick coat of mucin-like glycoproteins covers the surface of Trypanosoma cruzi and plays a crucial role in parasite protection and infectivity and host immunomodulation. The appealing candidate genes coding for the mucins of the mammal-dwelling stages define a heterogeneous family termed TcMUC, which comprises up to 700 members, thus precluding a genetic approach to address the protein core identity. Here, we demonstrate by multiple approaches that the TcMUC II genes code for the majority of trypomastigote mucins. These molecules display a variable, non-repetitive, highly O-glycosylated central domain, followed by a short conserved C terminus and a glycosylphosphatidylinositol anchor. A simultaneous expression of multiple TcMUC II gene products was observed. Moreover, the C terminus of TcMUC II mucins, but not their central domain, elicited strong antibody responses in patients with Chagas' disease and T. crusi infected animals. This highly diverse coat of mucins may represent a refined parasite strategy to elude the mammalian host immune system.Trypanosoma cruzi is the etiologic agent of Chagas' disease, which is of major medical and economical significance in Latin America (1). The T. cruzi life cycle involves distinct stages in both the mammalian host and the hematophagous insect vector (2). Within the insect, two major developmental forms can be observed: replicative epimastigotes and metacyclic trypomastigotes. The latter form brings the infection into humans when released on the skin or mucosa after the insect blood meal. Following cell invasion, metacyclic trypomastigotes differentiate into amastigotes, which, after several divisions, transform into cell-derived trypomastigotes, which are then released into the bloodstream. This stage is able to invade a wide variety of nucleated cells, thus propagating the infection.A thick coat of glycoproteins covers the surface of all these developmental stages (3-11). The major protein components of this coat have been identified as glycosylphosphatidylinositol (GPI) 1 -anchored molecules enriched in Thr, Ser, and Pro residues that serve as a scaffold for the extensive addition of O-glycans (5, 8 -10, 12). This particular feature enables their classification as mucin-like proteins by analogy to mammalian mucins (13). Mucins play a key role in parasite protection and infectivity and modulation of the host immune response throughout the T. cruzi life cycle (14 -16). The mucin coat of the cell-derived trypomastigotes (tGPI-mucins) is composed of an undefined mixture of molecules ranging from 60 to 220 kDa (6, 7, 9) and sharing the stage-specific, sialic acid-containing epitope Ssp-3, critical for mammalian cell attachment/invasion (17, 18). tGPI-mucins or their GPI moieties are potent inducers of nitric oxide and pro-inflammatory cytokines by macrophages (15,19). Major protective lytic antibodies directed against ␣-galactosyl epitopes present in tGPI-mucins have been described in sera from chronic Chagas' disease patients (6, 7, 9, 11).Recently, substantial informatio...

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Trypanosoma cruzi genome project: biological characteristics and molecular typing of clone CL Brener

Cited by 75 publications

References 25 publications

The autophagic pathway is a key component in the lysosomal dependent entry ofTrypanosoma cruziinto the host cell

The autophagic pathway is a key component in the lysosomal dependent entry ofTrypanosoma cruziinto the host cell

mRNA maturation by two-step trans-splicing/polyadenylation processing in trypanosomes

The Surface Coat of the Mammal-dwelling Infective Trypomastigote Stage of Trypanosoma cruzi Is Formed by Highly Diverse Immunogenic Mucins

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