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Infection and Immunity, July 2003, p. 4011-4017, Vol. 71, No. 7
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.7.4011-4017.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

The Delta Subunit of RNA Polymerase Is Required for Virulence of Streptococcus agalactiae

Amanda L. Jones,* Rachel H. V. Needham, and Craig E. Rubens

Department of Pediatrics, Division of Infectious Diseases, Children's Hospital and Regional Medical Center and University of Washington, Seattle, Washington 98105

Received 15 January 2003/ Returned for modification 12 March 2003/ Accepted 27 March 2003

Group B streptococci (GBS) remain the most significant bacterial pathogen causing neonatal sepsis, pneumonia, and meningitis in the United States despite the chemoprophylaxis strategies for preventing infection recommended by the Centers for Disease Control and Prevention. Using signature-tagged transposon mutagenesis to screen for novel virulence factors, we identified the rpoE gene as essential for development of sepsis in a neonatal rat model of GBS infection. An rpoE allelic replacement mutant displayed attenuated virulence in the sepsis model of infection identical to that of the transposon mutant, confirming linkage of the phenotype to the mutation in rpoE. The rpoE mutants also displayed increased sensitivity to killing in whole-blood bactericidal assays, which may explain the attenuated virulence. The mutants were otherwise phenotypically identical to the wild-type strain, including growth rate in plasma, indicating that a growth defect is not responsible for the attenuated virulence. rpoE is found only in gram-positive bacterial species and encodes the delta peptide, a subunit of RNA polymerase. Previous in vitro studies in other bacteria suggest that the delta peptide plays a role in maintaining transcriptional fidelity by blocking RNA polymerase binding at all but the strongest promoters, thereby inhibiting initiation of transcription. Despite the availability of rpoE mutants for several gram-positive bacterial species, a role for the peptide in vivo has not been defined, though it has been postulated that the delta peptide may be important for long-term survival in vitro or during growth phase transitions. Our data represent the first report of a phenotype relevant to virulence for rpoE mutants.

* Corresponding author. Mailing address: Department of Pediatrics, Division of Infectious Diseases, Children's Hospital and Regional Medical Center, 4800 Sand Point Way N.E., Seattle, WA 98105. Phone: (206) 987-2075. Fax: (206) 987-3890. E-mail: amanda.jones{at}seattlechildrens.org.

Editor: J. N. Weiser

Infection and Immunity, July 2003, p. 4011-4017, Vol. 71, No. 7
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.7.4011-4017.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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