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Comparative Proteomic Analysis of the Mycobacterium tuberculosis Lineage 7 Reveals Differentially Abundant Proteins Linked to Slow Growth

ABSTRACT: Differentially abundant proteins were quantified in the slowly growing Mycobacterium tuberculosis (M. tuberculosis) lineage 7 strains and the M. tuberculosis lineage 4 reference strain H37Rv. Mass spectrometry based proteomic analysis was employed to identify, quantitate and compare the protein profiles of strains from the two M. tuberculosis lineages (lineage 7 and 4). Label-free peptide quantification of whole cells from M. tuberculosis lineage 7 and 4 yielded the identification of 2825 and 2541 proteins, respectively. A combined total of 2867 protein groups covering 71% of the predicted M. tuberculosis proteome were identified. Comparative proteomic mapping of M. tuberculosis lineage 7 and 4 strains showed that the abundance of 125 proteins was significantly altered. Functional analysis showed that a number of M. tuberculosis proteins involved in growth and virulence were less abundant in lineage 7 compared to lineage 4. Five ABC transporter proteins, three phosphate binding proteins essential for inorganic phosphate uptake, and six components of the type 7 secretion system ESX-3 involved in iron acquisition were less abundant in M. tuberculosis lineage 7. This proteogenomic analysis provided an insight into the lineage 7-specific protein profile which may provide cues to understanding the differential properties of lineage 7 strains in terms of slow growth, survival fitness and pathogenesis.

INSTRUMENT(S):

ORGANISM(S): Mycobacterium Tuberculosis H37rv

TISSUE(S): Cell Lysate

DISEASE(S): Tuberculosis

SUBMITTER: Shewit Kalayou

LAB HEAD: Prof Tone Tønjum

PROVIDER: PXD006117 | Pride | 2017-05-24

REPOSITORIES: Pride

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Comparative Proteomic Analysis of Mycobacterium tuberculosis Lineage 7 and Lineage 4 Strains Reveals Differentially Abundant Proteins Linked to Slow Growth and Virulence.

Yimer Solomon A SA Birhanu Alemayehu G AG Kalayou Shewit S Riaz Tahira T Zegeye Ephrem D ED Beyene Getachew T GT Holm-Hansen Carol C Norheim Gunnstein G Abebe Markos M Aseffa Abraham A Tønjum Tone T

Frontiers in microbiology 20170509

In order to decipher the nature of the slowly growing Mycobacterium tuberculosis (M.tuberculosis) lineage 7, the differentially abundant proteins in strains of M. tuberculosis lineage 7 and lineage 4 were defined. Comparative proteomic analysis by mass spectrometry was employed to identify, quantitate and compare the protein profiles of strains from the two M. tuberculosis lineages. Label-free peptide quantification of whole cells from M. tuberculosis lineage 7 and 4 ...[more]

PMID: 28536560

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Project description:Influenza B virus (IBV) strains are one of the components of seasonal influenza vaccines in both trivalent and quadrivalent formulations. The vast majority of these vaccines are produced in embryonated chickens' eggs. While optimized backbones for vaccine production in eggs exist and are in use for influenza A viruses, no such backbones exist for IBVs, resulting in unpredictable production yields. To generate an optimal vaccine seed virus backbone, we have compiled a panel of 71 IBV strains from 1940 to present day, representing the known temporal and genetic variability of IBV circulating in humans. This panel contains strains from the B/Victoria/2/87-like lineage, B/Yamagata/16/88-like lineage and the ancestral lineage that preceded their split to provide a diverse set that would help to identify a suitable backbone which can be used in combination with hemagglutinin (HA) and neuraminidase (NA) glycoproteins from any IBV strain to be incorporated into the seasonal vaccine. We have characterized and ranked the growth profiles of the 71 IBV strains and the best performing strains were used for co-infection of eggs, followed by serial passaging to select for high-growth reassortant viruses. After serial passaging, we selected 10 clonal isolates based on their growth profiles assessed by hemagglutination and plaque-forming units. We then generated reverse genetics systems for the three clones that performed best in growth curves. The selected backbones were then used to generate different reassortant viruses with HA/NA combinations from high and low titer yielding wild type IBV. When the growth profiles of the recombinant reassortant viruses were tested, the low titer yielding HA/NA viruses with the selected backbones yielded higher titers similar to those from high titer yielding HA/NA combinations. The use of these IBV backbones with improved replication in eggs might increase yields for the influenza B virus components of seasonal influenza virus vaccines.

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Dataset Information

Comparative Proteomic Analysis of the Mycobacterium tuberculosis Lineage 7 Reveals Differentially Abundant Proteins Linked to Slow Growth

Dataset's files

Publications

Comparative Proteomic Analysis of <i>Mycobacterium tuberculosis</i> Lineage 7 and Lineage 4 Strains Reveals Differentially Abundant Proteins Linked to Slow Growth and Virulence.

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