Molecular characterization of the hexose transporter gene in benznidazole resistant and susceptible populations of Trypanosoma cruzi

Particle and Fibre Toxicology

Silvane M F Murta¹  Alvaro J Romanha¹  Guilherme Oliveira²  Edson L Folador¹  Antônio M Rezende¹  Douglas S Moreira¹  Rodrigo P P Soares¹  Jerônimo C Ruiz¹  Paula F dos Santos¹ 

[1] Centro de Pesquisas René Rachou/FIOCRUZ, Avenida Augusto de Lima 1715, Belo Horizonte, 30190-002, MG, Brazil;Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, 31270-910, MG, Brazil

关键词: Hexose transporters;    Drug resistance;    Trypanosoma cruzi;   

Others  :  1231727
DOI  :  10.1186/1756-3305-5-161

 received in 2012-03-26, accepted in 2012-08-02,  发布年份 2012

【 摘 要 】

Background

Hexose transporters (HT) are membrane proteins involved in the uptake of energy-supplying glucose and other hexoses into the cell. Previous studies employing the Differential Display technique have shown that the transcription level of the HT gene from T. cruzi (TcrHT) is higher in an in vitro-induced benznidazole (BZ)-resistant population of the parasite (17 LER) than in its susceptible counterpart (17 WTS).

Methods

In the present study, TcrHT has been characterized in populations and strains of T. cruzi that are resistant or susceptible to BZ. We investigated the copy number and chromosomal location of the gene, the levels of TcrHT mRNA and of TcrHT activity, and the phylogenetic relationship between TcrHT and HTs from other organisms.

Results

In silico analyses revealed that 15 sequences of the TcrHT gene are present in the T. cruzi genome, considering both CL Brener haplotypes. Southern blot analyses confirmed that the gene is present as a multicopy tandem array and indicated a nucleotide sequence polymorphism associated to T. cruzi group I or II. Karyotype analyses revealed that TcrHT is located in two chromosomal bands varying in size from 1.85 to 2.6 Mb depending on the strain of T. cruzi. The sequence of amino acids in the HT from T. cruzi is closely related to the HT sequences of Leishmania species according to phylogenetic analysis. Northern blot and quantitative real-time reverse transcriptase polymerase chain reaction analyses revealed that TcrHT transcripts are 2.6-fold higher in the resistant 17 LER population than in the susceptible 17 WTS. Interestingly, the hexose transporter activity was 40% lower in the 17 LER population than in all other T. cruzi samples analyzed. This phenotype was detected only in the in vitro-induced BZ resistant population, but not in the in vivo-selected or naturally BZ resistant T. cruzi samples. Sequencing analysis revealed that the amino acid sequences of the TcrHT from 17WTS and 17LER populations are identical. This result suggests that the difference in glucose transport between 17WTS and 17LER populations is not due to point mutations, but probably due to lower protein expression level.

Conclusion

The BZ resistant population 17 LER presents a decrease in glucose uptake in response to drug pressure.

【 授权许可】

   
2012 dos Santos et al.; licensee BioMed Central Ltd.

【 图 表 】

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