Microarray analysis of pneumococcal gene expression during invasive disease - PubMed (original) (raw)

Microarray analysis of pneumococcal gene expression during invasive disease

Carlos J Orihuela et al. Infect Immun. 2004 Oct.

Abstract

Streptococcus pneumoniae is a leading cause of invasive bacterial disease. This is the first study to examine the expression of S. pneumoniae genes in vivo by using whole-genome microarrays available from The Institute for Genomic Research. Total RNA was collected from pneumococci isolated from infected blood, infected cerebrospinal fluid, and bacteria attached to a pharyngeal epithelial cell line in vitro. Microarray analysis of pneumococcal genes expressed in these models identified body site-specific patterns of expression for virulence factors, transporters, transcription factors, translation-associated proteins, metabolism, and genes with unknown function. Contributions to virulence predicted for several unknown genes with enhanced expression in vivo were confirmed by insertion duplication mutagenesis and challenge of mice with the mutants. Finally, we cross-referenced our results with previous studies that used signature-tagged mutagenesis and differential fluorescence induction to identify genes that are potentially required by a broad range of pneumococcal strains for invasive disease.

PubMed Disclaimer

Figures

FIG. 1.

FIG. 1.

RNA collected from S. pneumoniae in vivo and in vitro. After isolation of pneumococcal RNA from infected blood, CSF, and bacteria adherent to the Detroit pharyngeal epithelial cell line, RNA was visualized on a 1% TBE gel to confirm purity and assess degradation. Bacterial RNA (open arrowheads) and eukaryotic RNA (shaded arrowheads) are indicated. Bacterial RNA isolated from the ECC model was enriched after collection of the initial pellet (raw) to remove contaminating eukaryotic RNA.

FIG. 2.

FIG. 2.

Venn diagrams highlighting disparity between infectious models for genes with altered expression. Numbers indicate the amount of genes determined to have altered expression that are either shared or exclusive to growth of the pneumococci in blood, CSF, or ECC.

Similar articles

Cited by

References

    1. Allen, B. L., and M. Hook. 2002. Isolation of a putative laminin binding protein from Streptococcus anginosus. Microb. Pathog. 33:23-31. - PubMed
    1. Avery, O. T., C. M. MacLeod, and M. McCarty. 1944. Studies on the chemical nature of the substance inducing transformation of the pneumococcal types: induction of transformation by the deoxyribonucleic acid fraction isolated from pneumococcus type III. J. Exp. Med. 79:137-157. - PMC - PubMed
    1. Beam, C. E., C. J. Saveson, and S. T. Lovett. 2002. Role for radA/sms in recombination intermediate processing in Escherichia coli. J. Bacteriol. 184:6836-6844. - PMC - PubMed
    1. Bethe, G., R. Nau, A. Wellmer, R. Hakenbeck, R. R. Reinert, H. P. Heinz, and G. Zysk. 2001. The cell wall-associated serine protease PrtA: a highly conserved virulence factor of Streptococcus pneumoniae. FEMS Microbiol. Lett. 205:99-104. - PubMed
    1. Blue, C. E., G. K. Paterson, A. R. Kerr, M. Berge, J. P. Claverys, and T. J. Mitchell. 2003. ZmpB, a novel virulence factor of Streptococcus pneumoniae that induces tumor necrosis factor alpha production in the respiratory tract. Infect. Immun. 71:4925-4935. - PMC - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources