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Journal of Bacteriology, June 2005, p. 3969-3979, Vol. 187, No. 12
0021-9193/05/$08.00+0     doi:10.1128/JB.187.12.3969-3979.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Pneumococcal HtrA Protease Mediates Inhibition of Competence by the CiaRH Two-Component Signaling System

M. E. Sebert,1* K. P. Patel,1 M. Plotnick,2,{dagger} and J. N. Weiser1

Department of Microbiology,1 Pulmonary, Allergy and Critical Care Division, University of Pennsylvania, Philadelphia, Pennsylvania 191042

Received 19 November 2004/ Accepted 9 March 2005

Activation of the CiaRH two-component signaling system prevents the development of competence for genetic transformation in Streptococcus pneumoniae through a previously unknown mechanism. Earlier studies have shown that CiaRH controls the expression of htrA, which we show encodes a surface-expressed serine protease. We found that mutagenesis of the putative catalytic serine of HtrA, while not impacting the competence of a ciaRH+ strain, restored a normal competence profile to a strain having a mutation that constitutively activates the CiaH histidine kinase. This result implies that activity of HtrA is necessary for the CiaRH system to inhibit competence. Consistent with this finding, recombinant HtrA (rHtrA) decreased the competence of pneumococcal cultures. The rHtrA-mediated decline in transformation efficiency could not be corrected with excess competence-stimulating peptide (CSP), suggesting that HtrA does not act through degradation of this signaling molecule. The inhibitory effects of rHtrA and activated CiaH, however, were largely overcome in a strain having constitutive activation of the competence pathway through a mutation in the cytoplasmic domain of the ComD histidine kinase. Although these results suggested that HtrA might act through degradation of the extracellular portion of the ComD receptor, Western immunoblots for ComD did not reveal changes in protein levels attributable to HtrA. We therefore postulate that HtrA may act on an unknown protein target that potentiates the activation of the ComDE system by CSP. These findings suggest a novel regulatory role for pneumococcal HtrA in modulating the activity of a two-component signaling system that controls the development of genetic competence.

* Corresponding author. Mailing address: 402 Johnson Pavilion, 3610 Hamilton Walk, Philadelphia, PA 19104. Phone: (215) 573-3510. Fax: (215) 573-4856. E-mail: sebert{at}email.chop.edu.

{dagger} Present address: Centocor, Inc., Horsham, PA.

Journal of Bacteriology, June 2005, p. 3969-3979, Vol. 187, No. 12
0021-9193/05/$08.00+0     doi:10.1128/JB.187.12.3969-3979.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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