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The dUTPase Enzyme Is Essential in Mycobacterium smegmatis

Thymidine biosynthesis is essential in all cells. Inhibitors of the enzymes involved in this pathway (e.g. methotrexate) are thus frequently used as cytostatics. Due to its pivotal role in mycobacterial thymidylate synthesis dUTPase, which hydrolyzes dUTP into the dTTP precursor dUMP, has been sugge...

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Autores principales: Pecsi, Ildiko, Hirmondo, Rita, Brown, Amanda C., Lopata, Anna, Parish, Tanya, Vertessy, Beata G., Tóth, Judit
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3360063/
https://www.ncbi.nlm.nih.gov/pubmed/22655049
http://dx.doi.org/10.1371/journal.pone.0037461
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author Pecsi, Ildiko
Hirmondo, Rita
Brown, Amanda C.
Lopata, Anna
Parish, Tanya
Vertessy, Beata G.
Tóth, Judit
author_facet Pecsi, Ildiko
Hirmondo, Rita
Brown, Amanda C.
Lopata, Anna
Parish, Tanya
Vertessy, Beata G.
Tóth, Judit
author_sort Pecsi, Ildiko
collection PubMed
description Thymidine biosynthesis is essential in all cells. Inhibitors of the enzymes involved in this pathway (e.g. methotrexate) are thus frequently used as cytostatics. Due to its pivotal role in mycobacterial thymidylate synthesis dUTPase, which hydrolyzes dUTP into the dTTP precursor dUMP, has been suggested as a target for new antitubercular agents. All mycobacterial genomes encode dUTPase with a mycobacteria-specific surface loop absent in the human dUTPase. Using Mycobacterium smegmatis as a fast growing model for Mycobacterium tuberculosis, we demonstrate that dUTPase knock-out results in lethality that can be reverted by complementation with wild-type dUTPase. Interestingly, a mutant dUTPase gene lacking the genus-specific loop was unable to complement the knock-out phenotype. We also show that deletion of the mycobacteria-specific loop has no major effect on dUTPase enzymatic properties in vitro and thus a yet to be identified loop-specific function seems to be essential within the bacterial cell context. In addition, here we demonstrated that Mycobacterium tuberculosis dUTPase is fully functional in Mycobacterium smegmatis as it rescues the lethal knock-out phenotype. Our results indicate the potential of dUTPase as a target for antitubercular drugs and identify a genus-specific surface loop on the enzyme as a selective target.
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spelling pubmed-33600632012-05-31 The dUTPase Enzyme Is Essential in Mycobacterium smegmatis Pecsi, Ildiko Hirmondo, Rita Brown, Amanda C. Lopata, Anna Parish, Tanya Vertessy, Beata G. Tóth, Judit PLoS One Research Article Thymidine biosynthesis is essential in all cells. Inhibitors of the enzymes involved in this pathway (e.g. methotrexate) are thus frequently used as cytostatics. Due to its pivotal role in mycobacterial thymidylate synthesis dUTPase, which hydrolyzes dUTP into the dTTP precursor dUMP, has been suggested as a target for new antitubercular agents. All mycobacterial genomes encode dUTPase with a mycobacteria-specific surface loop absent in the human dUTPase. Using Mycobacterium smegmatis as a fast growing model for Mycobacterium tuberculosis, we demonstrate that dUTPase knock-out results in lethality that can be reverted by complementation with wild-type dUTPase. Interestingly, a mutant dUTPase gene lacking the genus-specific loop was unable to complement the knock-out phenotype. We also show that deletion of the mycobacteria-specific loop has no major effect on dUTPase enzymatic properties in vitro and thus a yet to be identified loop-specific function seems to be essential within the bacterial cell context. In addition, here we demonstrated that Mycobacterium tuberculosis dUTPase is fully functional in Mycobacterium smegmatis as it rescues the lethal knock-out phenotype. Our results indicate the potential of dUTPase as a target for antitubercular drugs and identify a genus-specific surface loop on the enzyme as a selective target. Public Library of Science 2012-05-24 /pmc/articles/PMC3360063/ /pubmed/22655049 http://dx.doi.org/10.1371/journal.pone.0037461 Text en Pecsi et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Pecsi, Ildiko
Hirmondo, Rita
Brown, Amanda C.
Lopata, Anna
Parish, Tanya
Vertessy, Beata G.
Tóth, Judit
The dUTPase Enzyme Is Essential in Mycobacterium smegmatis
title The dUTPase Enzyme Is Essential in Mycobacterium smegmatis
title_full The dUTPase Enzyme Is Essential in Mycobacterium smegmatis
title_fullStr The dUTPase Enzyme Is Essential in Mycobacterium smegmatis
title_full_unstemmed The dUTPase Enzyme Is Essential in Mycobacterium smegmatis
title_short The dUTPase Enzyme Is Essential in Mycobacterium smegmatis
title_sort dutpase enzyme is essential in mycobacterium smegmatis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3360063/
https://www.ncbi.nlm.nih.gov/pubmed/22655049
http://dx.doi.org/10.1371/journal.pone.0037461
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