Cargando…
L-arginine availability and arginase activity: Characterization of amino acid permease 3 in Leishmania amazonensis
BACKGROUND: Leishmania uses the amino acid L-arginine as a substrate for arginase, enzyme that produces urea and ornithine, last precursor of polyamine pathway. This pathway is used by the parasite to replicate and it is essential to establish the infection in the mammalian host. L-arginine is not s...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5693463/ https://www.ncbi.nlm.nih.gov/pubmed/29073150 http://dx.doi.org/10.1371/journal.pntd.0006025 |
_version_ | 1783279944710750208 |
---|---|
author | Aoki, Juliana Ide Muxel, Sandra Marcia Zampieri, Ricardo Andrade Acuña, Stephanie Maia Fernandes, Juliane Cristina Ribeiro Vanderlinde, Rubia Heloisa Sales, Maria Carmen Oliveira de Pinho Floeter-Winter, Lucile Maria |
author_facet | Aoki, Juliana Ide Muxel, Sandra Marcia Zampieri, Ricardo Andrade Acuña, Stephanie Maia Fernandes, Juliane Cristina Ribeiro Vanderlinde, Rubia Heloisa Sales, Maria Carmen Oliveira de Pinho Floeter-Winter, Lucile Maria |
author_sort | Aoki, Juliana Ide |
collection | PubMed |
description | BACKGROUND: Leishmania uses the amino acid L-arginine as a substrate for arginase, enzyme that produces urea and ornithine, last precursor of polyamine pathway. This pathway is used by the parasite to replicate and it is essential to establish the infection in the mammalian host. L-arginine is not synthesized by the parasite, so its uptake occurs through the amino acid permease 3 (AAP3). AAP3 is codified by two copies genes (5.1 and 4.7 copies), organized in tandem in the parasite genome. One copy presents the expression regulated by L-arginine availability. METHODOLOGY/PRINCIPAL FINDINGS: RNA-seq data revealed 14 amino acid transporters differentially expressed in the comparison of La-WT vs. La-arg(-) promastigotes and axenic amastigotes. The 5.1 and 4.7 aap3 transcripts were down-regulated in La-WT promastigotes vs. axenic amastigotes, and in La-WT vs. La-arg(-) promastigotes. In contrast, transcripts of other transporters were up-regulated in the same comparisons. The amount of 5.1 and 4.7 aap3 mRNA of intracellular amastigotes was also determined in sample preparations from macrophages, obtained from BALB/c and C57BL/6 mice and the human THP-1 lineage infected with La-WT or La-arg(-), revealing that the genetic host background is also important. We also determined the aap3 mRNA and AAP3 protein amounts of promastigotes and axenic amastigotes in different environmental growth conditions, varying pH, temperature and L-arginine availability. Interestingly, the increase of temperature increased the AAP3 level in plasma membrane and consequently the L-arginine uptake, independently of pH and L-arginine availability. In addition, we demonstrated that besides the plasma membrane localization, AAP3 was also localized in the glycosome of L. amazonensis promastigotes and axenic amastigotes. CONCLUSIONS/SIGNIFICANCE: In this report, we described the differential transcriptional profiling of amino acids transporters from La-WT and La-arg(-) promastigotes and axenic amastigotes. We also showed the increased AAP3 levels under amino acid starvation or its decrease in L-arginine supplementation. The differential AAP3 expression was determined in the differentiation of promastigotes to amastigotes conditions, as well as the detection of AAP3 in the plasma membrane reflecting in the L-arginine uptake. Our data suggest that depending on the amino acid pool and arginase activity, Leishmania senses and could use an alternative route for the amino acid transport in response to stress signaling. |
format | Online Article Text |
id | pubmed-5693463 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-56934632017-11-30 L-arginine availability and arginase activity: Characterization of amino acid permease 3 in Leishmania amazonensis Aoki, Juliana Ide Muxel, Sandra Marcia Zampieri, Ricardo Andrade Acuña, Stephanie Maia Fernandes, Juliane Cristina Ribeiro Vanderlinde, Rubia Heloisa Sales, Maria Carmen Oliveira de Pinho Floeter-Winter, Lucile Maria PLoS Negl Trop Dis Research Article BACKGROUND: Leishmania uses the amino acid L-arginine as a substrate for arginase, enzyme that produces urea and ornithine, last precursor of polyamine pathway. This pathway is used by the parasite to replicate and it is essential to establish the infection in the mammalian host. L-arginine is not synthesized by the parasite, so its uptake occurs through the amino acid permease 3 (AAP3). AAP3 is codified by two copies genes (5.1 and 4.7 copies), organized in tandem in the parasite genome. One copy presents the expression regulated by L-arginine availability. METHODOLOGY/PRINCIPAL FINDINGS: RNA-seq data revealed 14 amino acid transporters differentially expressed in the comparison of La-WT vs. La-arg(-) promastigotes and axenic amastigotes. The 5.1 and 4.7 aap3 transcripts were down-regulated in La-WT promastigotes vs. axenic amastigotes, and in La-WT vs. La-arg(-) promastigotes. In contrast, transcripts of other transporters were up-regulated in the same comparisons. The amount of 5.1 and 4.7 aap3 mRNA of intracellular amastigotes was also determined in sample preparations from macrophages, obtained from BALB/c and C57BL/6 mice and the human THP-1 lineage infected with La-WT or La-arg(-), revealing that the genetic host background is also important. We also determined the aap3 mRNA and AAP3 protein amounts of promastigotes and axenic amastigotes in different environmental growth conditions, varying pH, temperature and L-arginine availability. Interestingly, the increase of temperature increased the AAP3 level in plasma membrane and consequently the L-arginine uptake, independently of pH and L-arginine availability. In addition, we demonstrated that besides the plasma membrane localization, AAP3 was also localized in the glycosome of L. amazonensis promastigotes and axenic amastigotes. CONCLUSIONS/SIGNIFICANCE: In this report, we described the differential transcriptional profiling of amino acids transporters from La-WT and La-arg(-) promastigotes and axenic amastigotes. We also showed the increased AAP3 levels under amino acid starvation or its decrease in L-arginine supplementation. The differential AAP3 expression was determined in the differentiation of promastigotes to amastigotes conditions, as well as the detection of AAP3 in the plasma membrane reflecting in the L-arginine uptake. Our data suggest that depending on the amino acid pool and arginase activity, Leishmania senses and could use an alternative route for the amino acid transport in response to stress signaling. Public Library of Science 2017-10-26 /pmc/articles/PMC5693463/ /pubmed/29073150 http://dx.doi.org/10.1371/journal.pntd.0006025 Text en © 2017 Aoki 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 (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Aoki, Juliana Ide Muxel, Sandra Marcia Zampieri, Ricardo Andrade Acuña, Stephanie Maia Fernandes, Juliane Cristina Ribeiro Vanderlinde, Rubia Heloisa Sales, Maria Carmen Oliveira de Pinho Floeter-Winter, Lucile Maria L-arginine availability and arginase activity: Characterization of amino acid permease 3 in Leishmania amazonensis |
title | L-arginine availability and arginase activity: Characterization of amino acid permease 3 in Leishmania amazonensis |
title_full | L-arginine availability and arginase activity: Characterization of amino acid permease 3 in Leishmania amazonensis |
title_fullStr | L-arginine availability and arginase activity: Characterization of amino acid permease 3 in Leishmania amazonensis |
title_full_unstemmed | L-arginine availability and arginase activity: Characterization of amino acid permease 3 in Leishmania amazonensis |
title_short | L-arginine availability and arginase activity: Characterization of amino acid permease 3 in Leishmania amazonensis |
title_sort | l-arginine availability and arginase activity: characterization of amino acid permease 3 in leishmania amazonensis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5693463/ https://www.ncbi.nlm.nih.gov/pubmed/29073150 http://dx.doi.org/10.1371/journal.pntd.0006025 |
work_keys_str_mv | AT aokijulianaide larginineavailabilityandarginaseactivitycharacterizationofaminoacidpermease3inleishmaniaamazonensis AT muxelsandramarcia larginineavailabilityandarginaseactivitycharacterizationofaminoacidpermease3inleishmaniaamazonensis AT zampieriricardoandrade larginineavailabilityandarginaseactivitycharacterizationofaminoacidpermease3inleishmaniaamazonensis AT acunastephaniemaia larginineavailabilityandarginaseactivitycharacterizationofaminoacidpermease3inleishmaniaamazonensis AT fernandesjulianecristinaribeiro larginineavailabilityandarginaseactivitycharacterizationofaminoacidpermease3inleishmaniaamazonensis AT vanderlinderubiaheloisa larginineavailabilityandarginaseactivitycharacterizationofaminoacidpermease3inleishmaniaamazonensis AT salesmariacarmenoliveiradepinho larginineavailabilityandarginaseactivitycharacterizationofaminoacidpermease3inleishmaniaamazonensis AT floeterwinterlucilemaria larginineavailabilityandarginaseactivitycharacterizationofaminoacidpermease3inleishmaniaamazonensis |