Selective amplification of maxicircle classes during the life cycle of Leishmania major

Flegontov, Pavel; Zhirenkina, E N; Gerasimov, Evgeny S.; Ponirovsky, E. N.; Strelkova, M. V.; Kolesnikov, Alexander

doi:10.1016/j.molbiopara.2009.01.014

Cited by 10 publications

(4 citation statements)

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“…Mutation in length or nucleotide composition and/or bi-parental inheritance in genetic exchange events are both exacerbated by differential replication rates and inequitable cytoplasmic segregation of mitochondrial genomes during mitosis [50], [51]. In kinetoplastids, maxicircle intra-clone diversity in the non-coding region was previously reported in both T. cruzi [31] and Leishmania major [52], [53]. In addition, an earlier study attributed a change in T. cruzi maxicircle gene repertoire (elimination of one of two heteroplasmic ND7 amplicons) to sub-culture [54].…”

Section: Discussionmentioning

confidence: 95%

Multiple Mitochondrial Introgression Events and Heteroplasmy in Trypanosoma cruzi Revealed by Maxicircle MLST and Next Generation Sequencing

et al. 2012

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Background Mitochondrial DNA is a valuable taxonomic marker due to its relatively fast rate of evolution. In Trypanosoma cruzi , the causative agent of Chagas disease, the mitochondrial genome has a unique structural organization consisting of 20–50 maxicircles (∼20 kb) and thousands of minicircles (0.5–10 kb). T. cruzi is an early diverging protist displaying remarkable genetic heterogeneity and is recognized as a complex of six discrete typing units (DTUs). The majority of infected humans are asymptomatic for life while 30–35% develop potentially fatal cardiac and/or digestive syndromes. However, the relationship between specific clinical outcomes and T. cruzi genotype remains elusive. The availability of whole genome sequences has driven advances in high resolution genotyping techniques and re-invigorated interest in exploring the diversity present within the various DTUs. Methodology/Principal Findings To describe intra-DTU diversity, we developed a highly resolutive maxicircle multilocus sequence typing (mtMLST) scheme based on ten gene fragments. A panel of 32 TcI isolates was genotyped using the mtMLST scheme, GPI , mini-exon and 25 microsatellite loci. Comparison of nuclear and mitochondrial data revealed clearly incongruent phylogenetic histories among different geographical populations as well as major DTUs. In parallel, we exploited read depth data, generated by Illumina sequencing of the maxicircle genome from the TcI reference strain Sylvio X10/1, to provide the first evidence of mitochondrial heteroplasmy (heterogeneous mitochondrial genomes in an individual cell) in T. cruzi . Conclusions/Significance mtMLST provides a powerful approach to genotyping at the sub-DTU level. This strategy will facilitate attempts to resolve phenotypic variation in T. cruzi and to address epidemiologically important hypotheses in conjunction with intensive spatio-temporal sampling. The observations of both general and specific incidences of nuclear-mitochondrial phylogenetic incongruence indicate that genetic recombination is geographically widespread and continues to influence the natural population structure of TcI, a conclusion which challenges the traditional paradigm of clonality in T. cruzi .

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Section: Discussionmentioning

confidence: 95%

Multiple Mitochondrial Introgression Events and Heteroplasmy in Trypanosoma cruzi Revealed by Maxicircle MLST and Next Generation Sequencing

et al. 2012

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“…However, other studies have shown evidence of the co-existence of mitochondrial mRNA populations with a different degree of editing in Leishmania spp. Additionally, maxicircle heteroplasmy was confirmed in similar studies analyzing the same strain of L. major , in which changes in the predominance maxicircle can take place during the differentiation of promastigote to amastigote state under stress conditions (Flegontov & Kolesnikov, 2006; Flegontov et al, 2009). Previously, it was reported that the ND8 in promastigotes of L. (V.) braziliensis must be edited to produce a functional transcript.…”

Section: Discussionmentioning

confidence: 52%

Mitochondrial genomics of human pathogenic parasiteLeishmania(Viannia)panamensis

Urrea

Triana-Chávez

Alzate

2019

PeerJ

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Background The human parasite Leishmania (V.) panamensis is one of the pathogenic species responsible for cutaneous leishmaniasis in Central and South America. Despite its importance in molecular parasitology, its mitochondrial genome, divided into minicircles and maxicircles, haven’t been described so far. Methods Using NGS-based sequencing (454 and ILLUMINA), and combining de novo genome assembly and mapping strategies, we report the maxicircle kDNA annotated genome of L. (V.) panamensis, the first reference of this molecule for the subgenus Viannia. A comparative genomics approach is performed against other Leishmania and Trypanosoma species. Results The results show synteny of mitochondrial genes of L. (V.) panamensis with other kinetoplastids. It was also possible to identify nucleotide variants within the coding regions of the maxicircle, shared among some of them and others specific to each strain. Furthermore, we compared the minicircles kDNA sequences of two strains and the results show that the conserved and divergent regions of the minicircles exhibit strain-specific associations.

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“…Whereas previous cases of imported leishmaniasis into Australia have been described (case studies 1-14) 29,30 , this study reports an additional 10 cases imported throughout 2017 (case studies [15][16][17][18][19][20][21][22][23][24]. Over the last two decades, a multitude of diagnostic assays targeting various genes including the ITS-1, Hsp70, antigen-encoding genes gp63 and cpb, RNA polymerase II, and the N-acetylglucosamine-1phosphate transferase have been designed for the detection and identification of Leishmania species.…”

Section: Discussionmentioning

confidence: 98%

“…20,21 It is composed of approximately 10,000 minicircles and 20-50 maxicircles that form the intricate network of the kinetoplast DNA (kDNA). [22][23][24][25] In a previous publication, we demonstrated the importance of the maxicircle for the taxonomic and phylogenetic analyses of Leishmania spp., providing a superior model to improve the resolution of trypanosomatid systematics. 26 Thus, using the advantageous properties of the maxicircle (i.e., potential to discriminate between closely related species) in the development, a quick and reliable test in the form of a PCR-RFLP has a clear potential in a laboratory setting.…”

Section: Introductionmentioning

confidence: 93%

Identification of Clinical Infections of Leishmania Imported into Australia: Revising Speciation with Polymerase Chain Reaction-RFLP of the Kinetoplast Maxicircle

Kaufer

Ellis

Stark

2019

The American Journal of Tropical Medicine and Hygiene

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Leishmaniasis is a vector-borne disease caused by protozoan parasites of the Leishmania genus. In Australia, leishmaniasis is an imported disease that is presenting itself at increased rates because of international travel, the influx of immigrants, and deployment of military operations to endemic regions. Although Leishmania species are morphologically indistinguishable, there is a strong correlation between some causative species of leishmaniasis and the subsequent response to the treatments available and patient outcome. Consequently, identification of the infective species is imperative as misidentification can result in the administering of an ineffective drug. The aim of this study was to develop a simple diagnostic tool with high sensitivity and specificity, which is capable of detecting the presence of the parasite and accurately differentiating the causative species in question. Using the advantageous properties of the maxi-circle kinetoplast DNA, a polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) targeting the ND7 gene was developed for the analysis of imported cases of human leishmaniasis in Australia. Designed as a dual analysis, concurrent PCR of Leishmania maxi-circle DNA and digestion with two separate enzymes (NlaIII and HpyCH4IV), this study provides an appraisal on 24 imported cases of leishmaniasis between 2008 and 2017. Five Leishmania species were reported, with members of the Viannia subgenus being the most common. The implementation of novel diagnostic procedures for leishmaniasis such as the one reported here is needed to establish a gold standard practice for the diagnosis and treatment of leishmaniasis.

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Selective amplification of maxicircle classes during the life cycle of Leishmania major

Cited by 10 publications

References 40 publications

Multiple Mitochondrial Introgression Events and Heteroplasmy in Trypanosoma cruzi Revealed by Maxicircle MLST and Next Generation Sequencing

Multiple Mitochondrial Introgression Events and Heteroplasmy in Trypanosoma cruzi Revealed by Maxicircle MLST and Next Generation Sequencing

Mitochondrial genomics of human pathogenic parasiteLeishmania(Viannia)panamensis

Identification of Clinical Infections of Leishmania Imported into Australia: Revising Speciation with Polymerase Chain Reaction-RFLP of the Kinetoplast Maxicircle

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