A functional genomic analysis of type 3 Streptococcus pneumoniae virulence

被引:232
作者
Lau, GW
Haataja, S
Lonetto, M
Kensit, SE
Marra, A
Bryant, AP
McDevitt, D
Morrison, DA
Holden, DW
机构
[1] Hammersmith Hosp, Imperial Coll Sch Med, Dept Infect Dis, London W12 0NN, England
[2] SmithKline Beecham PLC, Collegeville, PA 19426 USA
[3] Univ Illinois, Dept Biol Sci, Mol Biol Lab, Chicago, IL 60607 USA
关键词
D O I
10.1046/j.1365-2958.2001.02335.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Streptococcus pneumoniae remains a serious cause of morbidity and mortality in humans, but relatively little is known about the molecular basis of its pathogenesis. We used signature-tagged mutagenesis together with an analysis of S. pneumoniae genome sequence to identify and characterize genes required for pathogenesis. A library of signature-tagged mutants was created by insertion-duplication mutagenesis, and 1786 strains were analysed for their inability to survive and replicate in murine models of pneumonia and bacteraemia. One hundred and eighty-six mutant strains were identified as attenuated, and 56 were selected for further genetic characterization based on their ability to excise the integrated plasmid spontaneously. The genomic DNA inserts of the plasmids were cloned in Escherichia coli and sequenced. These sequences were subjected to database searches, including the S. pneumoniae genome sequence, which allowed us to examine the chromosomal regions flanking these genes. Most of the insertions were in probable operons, but no pathogenicity islands were found. Forty-two novel virulence loci were identified. Five strains mutated in genes involved in gene regulation, cation transport or stress tolerance were shown to be highly attenuated when tested individually in a murine respiratory tract infection model. Additional experiments also suggest that induction of competence for genetic transformation has a role in virulence.
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收藏
页码:555 / 571
页数:17
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