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Next generation sequencing of transcripts associated to five selected RNA-binding proteins in Leishmania infantum

ABSTRACT: Next generation sequencing of transcripts associated to five selected RNA-binding proteins in Leishmania infantum

PROVIDER: PRJNA675759 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13016025_1.fastq.gz	Fastqsanger.gz
	SRR13016025_2.fastq.gz	Fastqsanger.gz
	SRR13016026_1.fastq.gz	Fastqsanger.gz
	SRR13016026_2.fastq.gz	Fastqsanger.gz
	SRR13016027_1.fastq.gz	Fastqsanger.gz

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Next generation sequencing of transcripts associated to five selected RNA-binding proteins in Leishmania infantum

Project description:We report the sequencing of mRNAs associated with three Leishmania infantum Poly (A) binding proteins (PABP1, PABP2 and PABP3) and two of their partner RNA binding proteins (RBP23 and DRBD2). PABP1 and RBP23 preferentially associate with mRNAs that encode ribosomal proteins, absent from DRBD2-associated transcripts, and differently from PABP2 and PABP3 that bind to an overlapping set of mRNAs.

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Project description:The genomic DNAs of strains 263 of L. infantum and five derived independent resistant mutants to 5-fluorouracil were used in comparative genomic hybridizations to reveal the deletion and/or amplification events occured by drug resistance mechanisms. The human protozoan parasites Leishmania are prototrophic for pyrimidines and de novo pyrimidine biosynthesis is necessary for their growth. Five independent L. infantum mutants were selected for resistance to the pyrimidine analogue 5-fluorouracil (5-FU) in the hope to better understand the metabolism of pyrimidine in Leishmania. Analysis of the 5-FU mutants by comparative genomic hybridization and whole genome sequencing revealed amplification and deletion events as well as point mutations in metabolic genes involved in either the uridine salvage, folate or dTMP biosynthesis pathways. In particular, a dhfr-ts containing amplicon was observed in two mutants and a deletion of part of chromosome 10 was detected in one mutant. Point mutations in uridine phosphorybosyl transferase (UPRT), thymidine kinase (TK) and uridine phosphorylase (UP) were also discovered. Transfection experiments confirmed that these molecular alterations were responsible for the 5-FU resistance phenotype. Transport studies revealed that one resistant mutant was defective for uracil and 5-FU import although the identity of the transporter remains elusive. This study provided further insights in pyrimidine metabolism in Leishmania and confirmed that multiple mutations can co-exist in a cell to lead to resistance.

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