Novel DNA Binding and Regulatory Activities for σ 54 (RpoN) in Salmonella enterica Serovar Typhimurium 14028s

Author:

Bono Ashley C.1,Hartman Christine E.2,Solaimanpour Sina3,Tong Hao4,Porwollik Steffen5,McClelland Michael5,Frye Jonathan G.6,Mrázek Jan17,Karls Anna C.1

Affiliation:

1. Department of Microbiology, University of Georgia, Athens, Georgia, USA

2. Department of Genetics, University of Georgia, Athens, Georgia, USA

3. Department of Computer Science, University of Georgia, Athens, Georgia, USA

4. Department of Statistics, University of Georgia, Athens, Georgia, USA

5. Department of Microbiology and Molecular Genetics, University of California—Irvine School of Medicine, Irvine, California, USA

6. Bacterial Epidemiology and Antimicrobial Resistance Research Unit, U.S. Department of Agriculture, Athens, Georgia, USA

7. Institute of Bioinformatics, University of Georgia, Athens, Georgia, USA

Abstract

ABSTRACT The variable sigma (σ) subunit of the bacterial RNA polymerase (RNAP) holoenzyme, which is responsible for promoter specificity and open complex formation, plays a strategic role in the response to environmental changes. Salmonella enterica serovar Typhimurium utilizes the housekeeping σ 70 and five alternative sigma factors, including σ 54 . The σ 54 -RNAP differs from other σ-RNAP holoenzymes in that it forms a stable closed complex with the promoter and requires ATP hydrolysis by an activated cognate bacterial enhancer binding protein (bEBP) to transition to an open complex and initiate transcription. In S. Typhimurium, σ 54 -dependent promoters normally respond to one of 13 different bEBPs, each of which is activated under a specific growth condition. Here, we utilized a constitutively active, promiscuous bEBP to perform a genome-wide identification of σ 54 -RNAP DNA binding sites and the transcriptome of the σ 54 regulon of S. Typhimurium. The position and context of many of the identified σ 54 RNAP DNA binding sites suggest regulatory roles for σ 54 -RNAP that connect the σ 54 regulon to regulons of other σ factors to provide a dynamic response to rapidly changing environmental conditions. IMPORTANCE The alternative sigma factor σ 54 (RpoN) is required for expression of genes involved in processes with significance in agriculture, bioenergy production, bioremediation, and host-microbe interactions. The characterization of the σ 54 regulon of the versatile pathogen S. Typhimurium has expanded our understanding of the scope of the σ 54 regulon and how it links to other σ regulons within the complex regulatory network for gene expression in bacteria.

Funder

Binational Agriculture Research and Development Fund

North American Meat Institute

National Science Foundation

U.S. Department of Agriculture

Publisher

American Society for Microbiology

Subject

Molecular Biology,Microbiology

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