Transcriptomic profiling of gene expression and RNA processing during<i>Leishmania major</i>differentiation

Dillon, Laura A. L.; Okrah, Kwame; Hughitt, V. K.; Suresh, Rahul; Li, Yuan; Fernandes, Maria Cecília; Belew, Ashton T.; Bravo, Héctor Corrada; Mosser, David M.; El-Sayed, Najib M.

doi:10.1093/nar/gkv656

Cited by 80 publications

(109 citation statements)

References 92 publications

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“…Moreover, LmPRL-1 was still detectable in macrophage lysates even at 72 h after infection with promastigotes, when the parasites had already transformed into amastigotes. This observation on the protein level corroborates and extends recent mRNA expression profiling experiments (59,60). The expression of the gene LmjF.…”

Section: Discussionsupporting

confidence: 89%

Characterization of the Protein Tyrosine Phosphatase LmPRL-1 Secreted by Leishmania major via the Exosome Pathway

et al. 2017

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Similar to other intracellular pathogens, Leishmania parasites are known to evade the antimicrobial effector functions of host immune cells. To date, however, only a few virulence factors have been described for Leishmania major, one of the causative agents of cutaneous leishmaniasis. Here, we have characterized the expression and function of an L. major phosphatase, which we termed LmPRL-1. This enzyme shows a strong structural similarity to the human phosphatases of regenerating liver (PRL-1, -2, and -3) that regulate the proliferation, differentiation, and motility of cells. The biochemical characterization of the L. major phosphatase revealed that the enzyme is redox sensitive. When analyzing the subcellular localization of LmPRL-1 in promastigotes, amastigotes, and infected macrophages, we found that the phosphatase was predominantly expressed and secreted by promastigotes via the exosome route. Finally, we observed that ectopic expression of LmPRL-1 in L. major led to an increased number of parasites in macrophages. From these data, we conclude that the L. major phosphatase LmPRL-1 contributes to the intracellular survival of the parasites in macrophages.KEYWORDS Leishmania, exosome, macrophage, tyrosine phosphatase L eishmaniasis is an infectious disease prevalent worldwide that is caused by kinetoplastid protozoan parasites belonging to the genus Leishmania (1). In nature, this heteroxenous parasite is transmitted by the bites of sand flies. Following the inoculation of flagellated promastigotes into the dermis of the host, the parasites are rapidly endocytosed by phagocytic cells, in which they differentiate into amastigotes, multiply, and reach inner compartments and organs, such as draining lymph nodes, spleen, liver, and bone marrow. The life cycle of the parasite is completed after ingestion of amastigote-infected cells by sand flies during their blood meal. In the digestive tract of their vector, parasites transform back into extracellular promastigotes that develop into infectious metacyclics (2). Depending on the status of the host immune system and the Leishmania species, the infection of mammals leads either to mostly self-healing skin lesions (cutaneous leishmaniasis [CL]) or to a systemic disease, termed visceral leishmaniasis (VL) or kala-azar, that is lethal if untreated (3-5).To survive in their hosts, Leishmania parasites need to quickly adapt their growth, metabolism, and mechanisms of protection to their new environment. Interestingly, only a few genes are differentially expressed during this adaptive process, suggesting an important role for the regulation of protein translation (6). Leishmania speciesspecific gene diversity also participates in the adaptation of the parasite to its organ-

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Section: Discussionsupporting

confidence: 89%

Characterization of the Protein Tyrosine Phosphatase LmPRL-1 Secreted by Leishmania major via the Exosome Pathway

et al. 2017

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“…We compared the change in transcriptome between sand fly-derived PP and MP to the difference recently described between log-phase and metacyclic promastigotes in culture (7) (Tables 1 and 2). Six of the top 10 upregulated mRNAs observed in culture with metacyclics versus log-phase promastigotes were not significantly upregulated when sand fly-derived MP and PP were compared (Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…Importantly, whereas 6 out of the 10 most downregulated mRNAs in MP versus PP were the promastigote surface proteins (PSAs) on chromosome 12, these genes were not downregulated in cultured metacyclics, and in some cases they were slightly increased. Thirty-three GO categories were found to be significantly downregulated in metacyclics versus log-phase promastigotes in culture (see Table 2 in Dillon et al [7]). In contrast, only 8 GO categories were downregulated in MP versus PP in vivo , including those related to nucleosomes and ATP metabolism (Data Set S2).…”

Section: Resultsmentioning

confidence: 99%

“…The ability of the parasite to adapt to these vastly different environments has been the focus of a number of studies comparing the transcriptomes, proteomes, and metabolomes of amastigotes and promastigotes during their transformation in vitro (1–6). More recent studies have addressed the differential gene expression levels associated with the maturation of promastigotes to the mammalian-infective, metacyclic stage, again using in vitro conditions (7). Finally, two recent studies have demonstrated differential gene expression between L. infantum promastigtoes in the anterior midgut of a natural vector, Phlebotomus perniciosus , versus those in stationary-phase culture (8) or versus intracellular amastigotes in vitro (9).…”

Section: Introductionmentioning

confidence: 99%

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The Transcriptome ofLeishmania majorDevelopmental Stages in Their Natural Sand Fly Vector

Inbar

Hughitt

Dillon

et al. 2017

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148

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The life cycle of the Leishmania parasite in the sand fly vector involves differentiation into several distinctive forms, each thought to represent an adaptation to specific microenvironments in the midgut of the fly. Based on transcriptome sequencing (RNA-Seq) results, we describe the first high-resolution analysis of the transcriptome dynamics of four distinct stages of Leishmania major as they develop in a natural vector, Phlebotomus duboscqi. The early transformation from tissue amastigotes to procyclic promastigotes in the blood-fed midgut was accompanied by the greatest number of differentially expressed genes, including the downregulation of amastins, and upregulation of multiple cell surface proteins, sugar and amino acid transporters, and genes related to glucose metabolism and cell cycle progression. The global changes accompanying post-blood meal differentiation of procyclic promastigotes to the nectomonad and metacyclic stages were less extensive, though each displayed a unique signature. The transcriptome of nectomonads, which has not been studied previously, revealed changes consistent with cell cycle arrest and the upregulation of genes associated with starvation and stress, including autophagic pathways of protein recycling. Maturation to the infective, metacyclic stage was accompanied by changes suggesting preadaptation to the intracellular environment of the mammalian host, demonstrated by the amastigote-like profiles of surface proteins and metabolism-related genes. Finally, a direct comparison between sand fly-derived and culture-derived metacyclics revealed a reassuring similarity between the two forms, with the in vivo forms distinguished mainly by a stronger upregulation of transcripts associated with nutrient stress.

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“…Whole-transcriptome profiling revealed that calmodulin mRNA abundance in species of Leishmania is developmentally regulated. It is reduced by 1.5-2 fold upon differentiation from procyclics to metacyclics and amastigotes (Aslett et al 2010, Dillon et al 2015. Patterns of the expression of reporter genes upon differentiation of L. mexicana followed those of calmodulin.…”

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confidence: 92%

T7 polymerase-driven transcription is downregulated in metacyclic promastigotes and amastigotes of Leishmania mexicana

Ishemgulova¹,

Kraeva²,

Faktorová³

et al. 2016

FOLIA PARASIT

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Abstract:In our previous work we established a T7 polymerase-driven Tetracycline-inducible protein expression system in Leishmania mexicana (Biagi, 1953). We used this system to analyse gene expression profiles during development of L. mexicana in procyclic and metacyclic promastigotes and amastigotes. The transcription of the gene of interest and the T7 polymerase genes was significantly reduced upon cell differentiation. This regulation is not locus-specific. It depends on untranslated regions flanking open reading frames of the genes analysed. In this paper, we report that the previously established conventional inducible protein expression system may not be suitable for studies on differentiation of species of Leishmania Ross, 1903 and protein expression systems might have certain limitations.

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Transcriptomic profiling of gene expression and RNA processing duringLeishmania majordifferentiation

Cited by 80 publications

References 92 publications

Characterization of the Protein Tyrosine Phosphatase LmPRL-1 Secreted by Leishmania major via the Exosome Pathway

Characterization of the Protein Tyrosine Phosphatase LmPRL-1 Secreted by Leishmania major via the Exosome Pathway

The Transcriptome ofLeishmania majorDevelopmental Stages in Their Natural Sand Fly Vector

T7 polymerase-driven transcription is downregulated in metacyclic promastigotes and amastigotes of Leishmania mexicana

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