Bacteria
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA282954 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| PYXD01.dat.gz | Other | |||
| PYXD01.fasta.gz | Fasta.gz | |||
| PYXD01.master.dat | Other | |||
| PYXE01.dat.gz | Other | |||
| PYXE01.fasta.gz | Fasta.gz |
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Project description:Aggregatibacter actinomycetemcomitans (Aa) is a Gram-negative bacterial pathogen associated with severe periodontitis and non-oral diseases. Clinical isolates of Aa display a rough (R) colony phenotype with strong adherent properties. Upon prolonged culturing, non-adherent strains with a smooth (S) colony phenotype emerge. To date, most virulence studies on Aa have been performed with S strains of Aa, whereas the virulence of clinical R isolates received relatively little attention. Since the extracellular proteome is the main bacterial reservoir of virulence factors, the present study was aimed at a comparative analysis of this sub-proteome fraction for a collection of R isolates and derivative S strains, in order to link particular proteins to the virulence of Aa with serotype b. To assess the bacterial virulence, we applied different infection models based on larvae of the greater wax moth Galleria mellonella, a human salivary gland-derived epithelial cell line, and freshly isolated neutrophils from healthy human volunteers. A total number of 351 extracellular Aa proteins was identified by mass spectrometry, with the S strains consistently showing more extracellular proteins than their parental R isolates. A total of 50 known extracellular virulence factors was identified, of which 15 were expressed by all investigated bacteria. Importantly, the comparison of differences in exoproteome composition and virulence highlights critical roles of 10 extracellular proteins in the different infection models. Altogether, our present study provides novel cues for understanding the virulence of Aa, and for development of potential preventive or therapeutic avenues to neutralize this important oral pathogen.
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Project description:Six bacterial genomes, Geobacter metallireducens GS-15, Chromohalobacter salexigens, Vibrio breoganii 1C-10, Bacillus cereus ATCC 10987, Campylobacter jejuni subsp. jejuni 81-176 and Campylobacter jejuni NCTC 11168, all of which had previously been sequenced using other platforms were re-sequenced using single-molecule, real-time (SMRT) sequencing specifically to analyze their methylomes. In every case a number of new N6-methyladenine (m6A) and N4-methylcytosine (m4C) methylation patterns were discovered and the DNA methyltransferases (MTases) responsible for those methylation patterns were assigned. In 15 cases it was possible to match MTase genes with MTase recognition sequences without further sub-cloning. Two Type I restriction systems required sub-cloning to differentiate their recognition sequences, while four MTases genes that were not expressed in the native organism were sub-cloned to test for viability and recognition sequences. No attempt was made to detect 5-methylcytosine (m5C) recognition motifs from the SMRT sequencing data because this modification produces weaker signals using current methods. However, all predicted m6A and m4C MTases were detected unambiguously. This study shows that the addition of SMRT sequencing to traditional sequencing approaches gives a wealth of useful functional information about a genome showing not only which MTase genes are active, but also revealing their recognition sequences. Examination of the methylomes of six different strains of bacteria using kinetic data from single-molecule, real-time (SMRT) sequencing on the PacBio RS.
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