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Journal of Bacteriology, August 2001, p. 4876-4885, Vol. 183, No. 16
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.16.4876-4885.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

The HilA Box and Sequences outside It Determine the Magnitude of HilA-Dependent Activation of P_prgH from Salmonella Pathogenicity Island 1

C. Phoebe Lostroh and Catherine A. Lee^*

Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115

Received 23 March 2001/Accepted 1 June 2001

Salmonella requires genes on the Salmonella pathogenicity island 1 (SPI1) for the intestinal phase of infection in several models of pathogenesis. In Salmonella enterica serovar Typhimurium, most SPI1 genes are arranged in operons that are coordinately regulated by the SPI1-encoded protein HilA. In the past, it has been shown that HilA directly activates two promoters on SPI1, P_invF-1 and P_prgH. P_invF-1 contains a HilA binding site, termed a HilA box, that is necessary and sufficient for activation by HilA. The HilA box is 17 nucleotides long and contains a direct repeat comprised of two hexamers separated by 5 nucleotides, centered at 45 relative to the start site of transcription. P_prgH also contains a HilA box, and here we investigate its role at P_prgH. We have found that the HilA box is necessary, but not sufficient, for HilA-dependent activation of P_prgH. Instead, half-site-like hexamers outside the HilA box appear to be required for HilA-dependent activation of P_prgH, even though HilA binds to the HilA box in the absence of these hexamers. Thus, although HilA-dependent activation of P_invF-1 and P_prgH coordinates the expression of the structural genes for a type III secretion apparatus and the effectors secreted by that apparatus, it is also possible that mechanisms not apparent under in vitro inducing conditions could separate the expression of invFGEABC-spaMNOPQRS-sicA-sipBCDA-iacP-sicP-sptP and prgHIJK-orgABC.

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^* Corresponding author. Mailing address: Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Ave., Boston, MA 02115. Phone: (617) 432-4988. Fax: (617) 738-7664. E-mail: clee{at}hms.harvard.edu.

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Journal of Bacteriology, August 2001, p. 4876-4885, Vol. 183, No. 16
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.16.4876-4885.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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