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Infection and Immunity, July 2009, p. 2813-2823, Vol. 77, No. 7
0019-9567/09/$08.00+0 doi:10.1128/IAI.00060-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
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Department of Molecular Infectiology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chiba, Japan 260-8670
Received 16 January 2009/ Returned for modification 20 February 2009/ Accepted 10 April 2009
Shiga toxin 1 (Stx1) is located in the periplasmic fraction, while Stx2 is found in the extracellular fraction, suggesting that enterohemorrhagic Escherichia coli (EHEC) contains a specific Stx2 release system. Both stx1 and stx2 are found within the late operons of Stx-encoding phages. Stx2 production is greatly induced by mitomycin C, suggesting that stx2 can transcribe from the late phage promoter of the Stx2-encoding phage. However, the Stx1 promoter adjacent to stx1 is a dominant regulatory element in Stx1 production. With the deletion of phage lysis genes of the Stx2-encoding phage, Stx2 remains in the bacterial cells. Further, we demonstrate that the Stx2-encoding phage, but not the Stx1-encoding phage, is spontaneously induced at extremely low rates. These results indicate that spontaneously specific Stx2-encoding phage induction is involved in specific Stx2 release from bacterial cells. Furthermore, to examine whether another system for specific Stx2 release is present in EHEC, we analyze the stx-replaced mutants. As expected, Stx2 derived from the Stx1 promoter is located in both the extracellular and cell-associated fractions, while mutant Stx2 (B subunit, S31N) derived from the Stx1 promoter is found only in the cell-associated fraction. These results indicate that EHEC has another Stx2 release system that strictly recognizes the serine 31 residue of the B subunit. Overall, we present evidence that specific Stx2 release from bacterial cells is involved in both the Stx2-encoding phage induction system and another Stx2 release system.
Published ahead of print on 20 April 2009.
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