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Journal of Bacteriology, March 2009, p. 1498-1508, Vol. 191, No. 5
0021-9193/09/$08.00+0     doi:10.1128/JB.01177-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

T-POP Array Identifies EcnR and PefI-SrgD as Novel Regulators of Flagellar Gene Expression{triangledown} ,{dagger}

Christopher E. Wozniak,1,{ddagger} Changhan Lee,1 and Kelly T. Hughes2*

Department of Microbiology, University of Washington, Seattle, Washington 98195,1 Department of Biology, University of Utah, Salt Lake City, Utah 841122

Received 20 August 2008/ Accepted 15 December 2008

The T-POP transposon was employed in a general screen for tetracycline (Tet)-induced chromosomal loci that exhibited Tet-activated or Tet-repressed expression of a fliC-lac transcriptional fusion. Insertions that activated flagellar transcription were located in flagellar genes. T-POP insertions that exhibited Tet-dependent fliC-lac inhibition were isolated upstream of the ecnR, fimZ, pefI-srgD, rcsB, and ydiV genes and in the flagellar gene flgA, which is located upstream of the anti-{sigma}28 factor gene flgM. When expressed from the chromosomal ParaBAD promoter, EcnR, FimZ, PefI-SrgD, and RcsB inhibited the transcription of the flagellar class 1 flhDC operon. YdiV, which is weakly homologous to EAL domain proteins involved in cyclic-di-GMP regulation, appears to act at a step after class 1 transcription. By using a series of deletions of the regulatory genes to try to disrupt each pathway, these regulators were found to act largely independently of one another. These results identify EcnR and PefI-SrgD as additional components of the complex regulatory network controlling flagellar expression.

* Corresponding author. Mailing address: Department of Biology, University of Utah, 257 S. 1400 E., Salt Lake City, UT 84112-0840. Phone: (801) 585-6950. Fax: (801) 585-9735. E-mail: hughes{at}biology.utah.edu

{triangledown} Published ahead of print on 29 December 2008.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

{ddagger} Present address: Department of Biology, University of Utah, Salt Lake City, UT 84112.

Journal of Bacteriology, March 2009, p. 1498-1508, Vol. 191, No. 5
0021-9193/09/$08.00+0     doi:10.1128/JB.01177-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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