Genomewide Analysis of Mode of Action of the <italic toggle="yes">S</italic>-Adenosylmethionine Analogue Sinefungin in <named-content content-type="genus-species">Leishmania infantum</named-content>

ABSTRACT To further our understanding of one-carbon metabolism in the protozoan parasite Leishmania, we conducted genomic screens to study how the parasite responded to sinefungin (SNF) selection. SNF is a structural analogue of S-adenosylmethionine (AdoMet), a key methyl group donor to a number of...

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Autores principales:	Arijit Bhattacharya, Mansi Sharma, Charles Packianathan, Barry P. Rosen, Philippe Leprohon, Marc Ouellette
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Leishmania single-nucleotide variants copy number variation transporter methyltransferase S-adenosylmethionine Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/5a99ca2c131d4fb080e222bd46081b2d
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spelling	oai:doaj.org-article:5a99ca2c131d4fb080e222bd46081b2d2021-12-02T19:47:38ZGenomewide Analysis of Mode of Action of the <italic toggle="yes">S</italic>-Adenosylmethionine Analogue Sinefungin in <named-content content-type="genus-species">Leishmania infantum</named-content>10.1128/mSystems.00416-192379-5077https://doaj.org/article/5a99ca2c131d4fb080e222bd46081b2d2019-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSystems.00416-19https://doaj.org/toc/2379-5077ABSTRACT To further our understanding of one-carbon metabolism in the protozoan parasite Leishmania, we conducted genomic screens to study how the parasite responded to sinefungin (SNF) selection. SNF is a structural analogue of S-adenosylmethionine (AdoMet), a key methyl group donor to a number of biomolecules. One screen consisted of sequencing SNF-resistant mutants generated by stepwise selection with gradually increasing drug concentrations. These studies demonstrated deletion of the AdoMet transporter (AdoMetT1) by intergenic recombination as a crucial loss-of-function marker for SNF resistance. The second screen consisted of Cos-seq, a gain-of-function cosmid-based genomewide functional screen with increasing SNF concentration coupled to next-generation sequencing. Cosmids enriched in that screen and sequenced led to the identification of (i) the AdoMet synthetase (METK) as the major SNF target, (ii) an mRNA [(guanine-N7)-methyltransferase (CMT1)], (iii) a leucine carboxyl methyltransferase (LCMT), (iv) two tryparedoxin genes, and (v) two protein phosphatase regulatory genes. Further functional exploration indicated that LCMT interacts with one phosphatase catalytic subunit (PP2AC) and that mutation of the C-terminal leucine residue of PP2AC affects sinefungin susceptibility. These holistic screens led to the identification of transporters, biosynthetic genes, RNA and protein methyltransferases, as well as phosphatases linked to AdoMet-mediated functions in Leishmania. IMPORTANCE The two main cellular metabolic one-carbon donors are reduced folates and S-adenosylmethionine, whose biosynthetic pathways have proven highly effective in chemotherapeutic interventions in various cell types. Sinefungin, a nucleoside analogue of S-adenosylmethionine, was shown to have potent activity against the protozoan parasite Leishmania. Here, we studied resistance to sinefungin using whole-genome approaches as a way to further our understanding of the role of S-adenosylmethionine in this parasite and to reveal novel potential drug targets. These approaches allowed the characterization of novel features related to S-adenosylmethionine function in Leishmania which could further help in the development of sinefungin-like compounds against this pathogenic parasite.Arijit BhattacharyaMansi SharmaCharles PackianathanBarry P. RosenPhilippe LeprohonMarc OuelletteAmerican Society for MicrobiologyarticleLeishmaniasingle-nucleotide variantscopy number variationtransportermethyltransferaseS-adenosylmethionineMicrobiologyQR1-502ENmSystems, Vol 4, Iss 5 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Leishmania single-nucleotide variants copy number variation transporter methyltransferase S-adenosylmethionine Microbiology QR1-502
spellingShingle	Leishmania single-nucleotide variants copy number variation transporter methyltransferase S-adenosylmethionine Microbiology QR1-502 Arijit Bhattacharya Mansi Sharma Charles Packianathan Barry P. Rosen Philippe Leprohon Marc Ouellette Genomewide Analysis of Mode of Action of the <italic toggle="yes">S</italic>-Adenosylmethionine Analogue Sinefungin in <named-content content-type="genus-species">Leishmania infantum</named-content>
description	ABSTRACT To further our understanding of one-carbon metabolism in the protozoan parasite Leishmania, we conducted genomic screens to study how the parasite responded to sinefungin (SNF) selection. SNF is a structural analogue of S-adenosylmethionine (AdoMet), a key methyl group donor to a number of biomolecules. One screen consisted of sequencing SNF-resistant mutants generated by stepwise selection with gradually increasing drug concentrations. These studies demonstrated deletion of the AdoMet transporter (AdoMetT1) by intergenic recombination as a crucial loss-of-function marker for SNF resistance. The second screen consisted of Cos-seq, a gain-of-function cosmid-based genomewide functional screen with increasing SNF concentration coupled to next-generation sequencing. Cosmids enriched in that screen and sequenced led to the identification of (i) the AdoMet synthetase (METK) as the major SNF target, (ii) an mRNA [(guanine-N7)-methyltransferase (CMT1)], (iii) a leucine carboxyl methyltransferase (LCMT), (iv) two tryparedoxin genes, and (v) two protein phosphatase regulatory genes. Further functional exploration indicated that LCMT interacts with one phosphatase catalytic subunit (PP2AC) and that mutation of the C-terminal leucine residue of PP2AC affects sinefungin susceptibility. These holistic screens led to the identification of transporters, biosynthetic genes, RNA and protein methyltransferases, as well as phosphatases linked to AdoMet-mediated functions in Leishmania. IMPORTANCE The two main cellular metabolic one-carbon donors are reduced folates and S-adenosylmethionine, whose biosynthetic pathways have proven highly effective in chemotherapeutic interventions in various cell types. Sinefungin, a nucleoside analogue of S-adenosylmethionine, was shown to have potent activity against the protozoan parasite Leishmania. Here, we studied resistance to sinefungin using whole-genome approaches as a way to further our understanding of the role of S-adenosylmethionine in this parasite and to reveal novel potential drug targets. These approaches allowed the characterization of novel features related to S-adenosylmethionine function in Leishmania which could further help in the development of sinefungin-like compounds against this pathogenic parasite.
format	article
author	Arijit Bhattacharya Mansi Sharma Charles Packianathan Barry P. Rosen Philippe Leprohon Marc Ouellette
author_facet	Arijit Bhattacharya Mansi Sharma Charles Packianathan Barry P. Rosen Philippe Leprohon Marc Ouellette
author_sort	Arijit Bhattacharya
title	Genomewide Analysis of Mode of Action of the <italic toggle="yes">S</italic>-Adenosylmethionine Analogue Sinefungin in <named-content content-type="genus-species">Leishmania infantum</named-content>
title_short	Genomewide Analysis of Mode of Action of the <italic toggle="yes">S</italic>-Adenosylmethionine Analogue Sinefungin in <named-content content-type="genus-species">Leishmania infantum</named-content>
title_full	Genomewide Analysis of Mode of Action of the <italic toggle="yes">S</italic>-Adenosylmethionine Analogue Sinefungin in <named-content content-type="genus-species">Leishmania infantum</named-content>
title_fullStr	Genomewide Analysis of Mode of Action of the <italic toggle="yes">S</italic>-Adenosylmethionine Analogue Sinefungin in <named-content content-type="genus-species">Leishmania infantum</named-content>
title_full_unstemmed	Genomewide Analysis of Mode of Action of the <italic toggle="yes">S</italic>-Adenosylmethionine Analogue Sinefungin in <named-content content-type="genus-species">Leishmania infantum</named-content>
title_sort	genomewide analysis of mode of action of the <italic toggle="yes">s</italic>-adenosylmethionine analogue sinefungin in <named-content content-type="genus-species">leishmania infantum</named-content>
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/5a99ca2c131d4fb080e222bd46081b2d
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Genomewide Analysis of Mode of Action of the <italic toggle="yes">S</italic>-Adenosylmethionine Analogue Sinefungin in <named-content content-type="genus-species">Leishmania infantum</named-content>

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