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Infection and Immunity, October 2004, p. 5582-5596, Vol. 72, No. 10
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.10.5582-5596.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Microarray Analysis of Pneumococcal Gene Expression during Invasive Disease

Carlos J. Orihuela,1 Jana N. Radin,1,2 Jack E. Sublett,1 Geli Gao,1 Deepak Kaushal,3 and Elaine I. Tuomanen1,2*

Department of Infectious Diseases,1 Hartwell Center for Bioinformatics and Biotechnology St. Jude Children's Research Hospital,3 Department of Molecular Sciences, University of Tennessee Health Science Center Memphis, Tennessee2

Received 23 March 2004/ Returned for modification 19 May 2004/ Accepted 29 June 2004

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.

* Corresponding author. Mailing address: St. Jude Children's Research Hospital, Mailstop 320, IRC 8057, 332 North Lauderdale Rd., Memphis, TN 38105. Phone: (901) 495-3486. Fax: (901) 495-3099. E-mail: elaine.tuomanen{at}stjude.org.

Editor: J. N. Weiser

Infection and Immunity, October 2004, p. 5582-5596, Vol. 72, No. 10
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.10.5582-5596.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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