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Proteome of a Moraxella catarrhalis Strain under Iron-Restricted Conditions.

ABSTRACT: Moraxella catarrhalis is a leading cause of otitis media and exacerbations of chronic obstructive pulmonary disease; however, its response to iron starvation during infection is not completely understood. Here, we announce a sequential window acquisition of all theoretical fragment ion spectra mass spectrometry (SWATH-MS) data set describing the differential expression of the M. catarrhalis CCRI-195ME proteome under iron-restricted versus iron-replete conditions.

SUBMITTER: Blakeway LV

PROVIDER: S-EPMC7082453 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Proteome of a Moraxella catarrhalis Strain under Iron-Restricted Conditions.

Blakeway Luke V LV Tan Aimee A Peak Ian R IR Atack John M JM Seib Kate L KL

Microbiology resource announcements 20200319 12

<i>Moraxella catarrhalis</i> is a leading cause of otitis media and exacerbations of chronic obstructive pulmonary disease; however, its response to iron starvation during infection is not completely understood. Here, we announce a sequential window acquisition of all theoretical fragment ion spectra mass spectrometry (SWATH-MS) data set describing the differential expression of the <i>M. catarrhalis</i> CCRI-195ME proteome under iron-restricted versus iron-replete conditions. ...[more]

PMID: 32193234

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SWATH-MS proteome of Moraxella catarrhalis under iron restricted conditions

Project description:Moraxella catarrhalis is a human-restricted bacterial pathogen that causes otitis media in children and exacerbations of chronic obstructive pulmonary disease in adults. Iron is a co-factor required for the function of proteins involved in respiration and DNA replication, and thus is essential to life for most bacterial species. As iron is sequestered by host tissues, all host-adapted pathogens encounter periods of iron starvation. Previous studies have defined the repertoire of genes required for growth of M. catarrhalis under iron restricted conditions, and described differential expression at the transcriptional level. However, global changes in protein expression in response to iron starvation have not been investigated. Here we provide a SWATH-MS dataset describing differential expression of the M. catarrhalis CCRI-195ME proteome under iron restricted versus iron replete conditions.

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Project description:Total RNA samples were isolated from M. catarrhalis grown in BHI (iron-replete condition, designated DF0) and in BHI containing 30 mM Desferal (iron-limiting condition, designated DF30). Total RNA and M. catarrhalis genome-directed primers (GDPs) were used for synthesis of Cy3- or Cy5-labeled cDNA samples. Three independent M. catarrhalis growth experiments and total RNA isolations were used for synthesis of cDNA samples, and a total of six DNA microarray hybridizations were performed in this study. Keywords: Effect of Growth Environment on Global Gene Expression by Moraxella catarrhalis

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Identification of genes essential for Moraxella catarrhalis growth under iron-limiting conditions

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Identification of genes essential for Moraxella catarrhalis growth under iron-limiting conditions using Genomic Array Footprinting (GAF)

Project description:Iron-sequestration by the human host is a first line defense against respiratory pathogens like Moraxella catarrhalis, which consequently experiences a period of iron-starvation during colonization and infection. We determined the genetic requirements for M. catarrhalis growth during iron-starvation using the high-throughput genome-wide screening technology genomic array footprinting (GAF). To this end, a large marinerT7 transposon mutant library (~28,000 independent transposon mutants) was grown under iron-limiting conditions, achieved by sequestration of iron by 30 µM Desferal (DF30), and under control growth conditions (brain heart infusion broth, DF0). Mutants were recovered at exponential- and the early-stationary growth phase and used for the generation of mutant-specific cDNA probes that were hybridized to custom-designed NimbleGen GAF microarrays. The results described in this study are further discussed in Stefan P.W. de Vries, Peter Burghout, Jeroen D. Langereis, Aldert Zomer, Peter W.M. Hermans, Hester J. Bootsma: Genetic requirements for Moraxella catarrhalis growth under iron-limiting conditions, Molecular Microbiology.

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Project description:Iron-sequestration by the human host is a first line defense against respiratory pathogens like Moraxella catarrhalis, which consequently experiences a period of iron-starvation during colonization and infection. We determined the genetic requirements for M. catarrhalis growth during iron-starvation using the high-throughput genome-wide screening technology genomic array footprinting (GAF). To this end, a large marinerT7 transposon mutant library (~28,000 independent transposon mutants) was grown under iron-limiting conditions, achieved by sequestration of iron by 30 M-BM-5M Desferal (DF30), and under control growth conditions (brain heart infusion broth, DF0). Mutants were recovered at exponential- and the early-stationary growth phase and used for the generation of mutant-specific cDNA probes that were hybridized to custom-designed NimbleGen GAF microarrays. The results described in this study are further discussed in Stefan P.W. de Vries, Peter Burghout, Jeroen D. Langereis, Aldert Zomer, Peter W.M. Hermans, Hester J. Bootsma: Genetic requirements for Moraxella catarrhalis growth under iron-limiting conditions, Molecular Microbiology. cDNA probes generated from mutants recovered during the exponential growth phase, or early-stationary phase under challenge (iron-limiting, DF30) or control conditions (DF0). Hybridization on GAF 4x72K custom design Nimblegen GAF arrays for read-out.

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