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1.
PLoS One ; 9(3): e89312, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24595068

RESUMEN

We present a systematic study of three bacterial species that belong to the class Mollicutes, the smallest and simplest bacteria, Spiroplasma melliferum, Mycoplasma gallisepticum, and Acholeplasma laidlawii. To understand the difference in the basic principles of metabolism regulation and adaptation to environmental conditions in the three species, we analyzed the metabolome of these bacteria. Metabolic pathways were reconstructed using the proteogenomic annotation data provided by our lab. The results of metabolome, proteome and genome profiling suggest a fundamental difference in the adaptation of the three closely related Mollicute species to stress conditions. As the transaldolase is not annotated in Mollicutes, we propose variants of the pentose phosphate pathway catalyzed by annotated enzymes for three species. For metabolite detection we employed high performance liquid chromatography coupled with mass spectrometry. We used liquid chromatography method - hydrophilic interaction chromatography with silica column - as it effectively separates highly polar cellular metabolites prior to their detection by mass spectrometer.


Asunto(s)
Metabolómica/métodos , Tenericutes/metabolismo , Vías Biosintéticas , Hidrolasas/metabolismo , Espectrometría de Masas , Metaboloma , Vía de Pentosa Fosfato , Terpenos/metabolismo
2.
PLoS One ; 8(2): e56577, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23437175

RESUMEN

Tuberculosis caused by multidrug-resistant (MDR) and extensively drug-resistant (XDR) Mycobacterium tuberculosis (MTB) strains is a growing problem in many countries. The availability of the complete nucleotide sequences of several MTB genomes allows to use the comparative genomics as a tool to study the relationships of strains and differences in their evolutionary history including acquisition of drug-resistance. In our work, we sequenced three genomes of Russian MTB strains of different phenotypes--drug susceptible, MDR and XDR. Of them, MDR and XDR strains were collected in Tomsk (Siberia, Russia) during the local TB outbreak in 1998-1999 and belonged to rare KQ and KY families in accordance with IS6110 typing, which are considered endemic for Russia. Based on phylogenetic analysis, our isolates belonged to different genetic families, Beijing, Ural and LAM, which made the direct comparison of their genomes impossible. For this reason we performed their comparison in the broader context of all M. tuberculosis genomes available in GenBank. The list of unique individual non-synonymous SNPs for each sequenced isolate was formed by comparison with all SNPs detected within the same phylogenetic group. For further functional analysis, all proteins with unique SNPs were ascribed to 20 different functional classes based on Clusters of Orthologous Groups (COG). We have confirmed drug resistant status of our isolates that harbored almost all known drug-resistance associated mutations. Unique SNPs of an XDR isolate CTRI-4(XDR), belonging to a Beijing family were compared in more detail with SNPs of additional 14 Russian XDR strains of the same family. Only type specific mutations in genes of repair, replication and recombination system (COG category L) were found common within this group. Probably the other unique SNPs discovered in CTRI-4(XDR) may have an important role in adaptation of this microorganism to its surrounding and in escape from antituberculosis drugs treatment.


Asunto(s)
Tuberculosis Extensivamente Resistente a Drogas/genética , Tuberculosis Extensivamente Resistente a Drogas/microbiología , Mycobacterium tuberculosis/clasificación , Mycobacterium tuberculosis/genética , Antituberculosos/farmacología , Hibridación Genómica Comparativa , ADN Bacteriano , Tuberculosis Extensivamente Resistente a Drogas/epidemiología , Genoma Bacteriano/efectos de los fármacos , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Pruebas de Sensibilidad Microbiana , Datos de Secuencia Molecular , Mycobacterium tuberculosis/aislamiento & purificación , Fenotipo , Filogenia , Federación de Rusia , Siberia
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