Schistosoma mansoni: Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination

Santos, Daniela Soares; Aguiar, Pedro Henrique Nascimento; Lobo, Francisco Pereira; Mourão, Marina Moraes; Tambor, José Humberto Machado; Valadão, Analina Furtado; Vilas-Boas, Adlane; Nóbrega, Francisco G.; LoVerde, Philip T.; Macedo, Andréa Mara; Pena, Sérgio Danilo Junho; Machado, Carlos Renato; Franco, Glória Regina

doi:10.1016/j.exppara.2007.02.012

Cited by 8 publications

(7 citation statements)

References 35 publications

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“…Regarding the S. japonicum genome, the number of enzymes involved in the NEDD8 conjugation process is the same as is found in S. mansoni, reinforcing the evolutive proximity between these species [35]. During this analysis the RBX protein was not found in the parasite EST database (http://www.genedb.org/genedb/smansoni/), despite the fact that this E3 ligase has been characterised in this helminth previously [36]. Furthermore, we have observed errors in the assembled genome sequence for a subunit of the E1 heterodimer (APP1), reinforcing the importance of re-annotating the putative proteins deposited in the genome database to increase data accuracy.…”

Section: Discussionmentioning

confidence: 63%

NEDD8 conjugation in Schistosoma mansoni: Genome analysis and expression profiles

Pereira

Gomes

Olmo

et al. 2013

Parasitology International

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NEDD8 is an ubiquitin-like molecule that covalently binds to target proteins through an enzymatic cascade analogous to ubiquitylation. This modifier is known to bind to p53 and p73, as well as all Cullin family proteins, which are essential components of Skp1/Cul-1/F-box protein (SCF)-like Ub ligase complexes. Here, we focused on a genomic analysis of the genes involved in the NEDD8 conjugation pathway in Schistosoma mansoni. The results revealed seven genes related to NEDD8 conjugation that are conserved in Schistosoma japonicum, Caenorhabditis elegans, Drosophila melanogaster and Homo sapiens. We performed quantitative RT-PCR (qRT-PCR), which showed differential profiles for Smnedd8, Smapp1, Smuba3, Smube2f, Smdcn1, Smrbx and Smsenp8 throughout the life cycle of S. mansoni. Upregulation was observed in 3-day-old schistosomula and adult worms for all analysed genes. We also analysed the transcription levels of Cullin family members Smp63 and Smp73, and observed upregulation in early schistosomula, while cercariae and adult worms showed expression levels similar to one another. Taken together, these results suggest that the NEDDylation/DeNEDDylation pathway controls important cellular regulators during worm development from cercariae to schistosomula and, finally, to adult.

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Section: Discussionmentioning

confidence: 63%

NEDD8 conjugation in Schistosoma mansoni: Genome analysis and expression profiles

Pereira

Gomes

Olmo

et al. 2013

Parasitology International

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“…By performing genome-wide drug sensitivity screens (chemogenomic profiling) [12], [13], [14], [15], [16], [17] of yeast mutants with the antimalarials quinine [18], St. John's Wort [19] and artemisinin [20], researchers were able to identify their primary targets as well as identify potential side effects. Furthermore, several groups have been able to complement yeast loss-of function mutations by expressing coding sequences from parasites such as Plasmodium [21], [22], Schistosoma [23], [24], [25], Leishmania [26] or Trypanosoma [6], [27], [28], [29], [30], [31].…”

Section: Introductionmentioning

confidence: 99%

Functional Expression of Parasite Drug Targets and Their Human Orthologs in Yeast

et al. 2011

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BackgroundThe exacting nutritional requirements and complicated life cycles of parasites mean that they are not always amenable to high-throughput drug screening using automated procedures. Therefore, we have engineered the yeast Saccharomyces cerevisiae to act as a surrogate for expressing anti-parasitic targets from a range of biomedically important pathogens, to facilitate the rapid identification of new therapeutic agents.Methodology/Principal FindingsUsing pyrimethamine/dihydrofolate reductase (DHFR) as a model parasite drug/drug target system, we explore the potential of engineered yeast strains (expressing DHFR enzymes from Plasmodium falciparum, P. vivax, Homo sapiens, Schistosoma mansoni, Leishmania major, Trypanosoma brucei and T. cruzi) to exhibit appropriate differential sensitivity to pyrimethamine. Here, we demonstrate that yeast strains (lacking the major drug efflux pump, Pdr5p) expressing yeast (ScDFR1), human (HsDHFR), Schistosoma (SmDHFR), and Trypanosoma (TbDHFR and TcDHFR) DHFRs are insensitive to pyrimethamine treatment, whereas yeast strains producing Plasmodium (PfDHFR and PvDHFR) DHFRs are hypersensitive. Reassuringly, yeast strains expressing field-verified, drug-resistant mutants of P. falciparum DHFR (Pfdhfr 51I,59R,108N) are completely insensitive to pyrimethamine, further validating our approach to drug screening. We further show the versatility of the approach by replacing yeast essential genes with other potential drug targets, namely phosphoglycerate kinases (PGKs) and N-myristoyl transferases (NMTs).Conclusions/SignificanceWe have generated a number of yeast strains that can be successfully harnessed for the rapid and selective identification of urgently needed anti-parasitic agents.

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“…Regarding IMPDH, this protein is responsible for the rate-limiting step in guanine nucleotide biosynthesis (Prosise and Luecke, 2003), and it has previously been identified in Schistosoma genome and transcriptome (Protasio et al, 2012). E3 ligase enzyme catalyzes protein ubiquitination, which regulates various biological processes through covalent modification of proteins and transcription factors, and ubiquitin is the most important protein of this process (Sun and Chen, 2004; Santos et al, 2007). It has been suggested that ubiquitination is of interest in S. mansoni because this process could be a potential target for the design of new drugs (Guerra-Sá et al, 2005), being ubiquitin protein ligase E3a a good target to be studied.…”

Section: Discussionmentioning

confidence: 99%

Comparative Proteomics Reveals Characteristic Proteins on Praziquantel-resistance inSchistosoma mansoni

Pinto-Almeida

Mendes

Ferreira³

et al. 2018

Preprint

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The extensive use of Praziquantel (PZQ), the only drug available to treat schistosomiasis, has brought concern about the emergence of PZQ-resistance/tolerance by Schistosoma spp., thus reaffirming an urge for the development of new treatment alternatives. Therefore, it is imperative and urgent to study this phenomenon trying to understand what is involved in its occurrence. Studies of Schistosoma spp. genome, transcriptome and proteome are crucial to better understand this situation. By stepwise drug pressure from a fully susceptible parasite strain, our group selected a S. mansoni variant strain stably resistant to PZQ and isogenic to its fully susceptible parental counterpart, except for the genetic determinants of PZQ-resistance phenotype. Based on this, the objective of this study was to compare the proteomes of both strains, identifying proteins from male and female adult worms of PZQ-resistant and PZQ-susceptible strains, exposed and not exposed to PZQ, which were separated by high-resolution two-dimensional electrophoresis and sequenced by high throughput LC-MS/MS. Likewise, this work is extremely relevant since for the first time the proteome of a S. mansoni PZQ-resistant strain is studied and compared to the proteome of the respective S. mansoni PZQ-susceptible strain. This study identified 60 S. mansoni proteins, some of which differentially expressed in either strain, which may putatively be involved in the PZQ-resistance phenomenon. This information represents substantial progress towards deciphering the worm proteome. Furthermore, these data may constitute an informative source for further investigations into PZQ-resistance and increase the possibility of identifying proteins related to this condition, possibly contributing to avoid or decrease the likelihood of development and spread of PZQ-resistance. This is an innovative study that opens doors to PZQ-resistance surveys, contributing to discover a solution to PZQ-resistance problem, as suggests new potential targets for study.

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Schistosoma mansoni: Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination

Cited by 8 publications

References 35 publications

NEDD8 conjugation in Schistosoma mansoni: Genome analysis and expression profiles

NEDD8 conjugation in Schistosoma mansoni: Genome analysis and expression profiles

Functional Expression of Parasite Drug Targets and Their Human Orthologs in Yeast

Comparative Proteomics Reveals Characteristic Proteins on Praziquantel-resistance inSchistosoma mansoni

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