Role of acetyl-phosphate in activation of the Rrp2-RpoN-RpoS pathway in borrelia burgdorferi

Haijun Xu, Melissa J. Caimano, Tao Lin, Ming He, Justin D. Radolf, Steven J. Norris, Frank Gheradini, Alan J. Wolfe, X. Yang

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Borrelia burgdorferi, the Lyme disease spirochete, dramatically alters its transcriptome and proteome as it cycles between the arthropod vector and mammalian host. During this enzootic cycle, a novel regulatory network, the Rrp2-RpoN-RpoS pathway (also known as the σ54S sigma factor cascade), plays a central role in modulating the differential expression of more than 10% of all B. burgdorferi genes, including the major virulence genes ospA and ospC. However, the mechanism(s) by which the upstream activator and response regulator Rrp2 is activated remains unclear. Here, we show that none of the histidine kinases present in the B. burgdorferi genome are required for the activation of Rrp2. Instead, we present biochemical and genetic evidence that supports the hypothesis that activation of the Rrp2-RpoN-RpoS pathway occurs via the small, high-energy, phosphoryl-donor acetyl phosphate (acetyl~P), the intermediate of the Ack-Pta (acetate kinase-phosphate acetyltransferase) pathway that converts acetate to acetyl-CoA. Supplementation of the growth medium with acetate induced activation of the Rrp2-RpoN-RpoS pathway in a dose-dependent manner. Conversely, the overexpression of Pta virtually abolished acetate-induced activation of this pathway, suggesting that acetate works through acetyl,P. Overexpression of Pta also greatly inhibited temperature and cell density-induced activation of RpoS and OspC, suggesting that these environmental cues affect the Rrp2-RpoN-RpoS pathway by influencing acetyl~P. Finally, overexpression of Pta partially reduced infectivity of B. burgdorferi in mice. Taken together, these findings suggest that acetyl~P is one of the key activating molecule for the activation of the Rrp2-RpoN-RpoS pathway and support the emerging concept that acetyl~P can serve as a global signal in bacterial pathogenesis.

Original languageEnglish
Article numbere01104
JournalPLoS Pathogens
Volume6
Issue number9
DOIs
StatePublished - Sep 2010

Fingerprint

Borrelia burgdorferi
Acetates
RNA Polymerase Sigma 54
Phosphate Acetyltransferase
Acetate Kinase
Arthropod Vectors
Acetyl Coenzyme A
Proteome
Transcriptome
Genes
Cues
Virulence
Molecular Biology
Cell Count
acetyl phosphate
Genome
Temperature
Growth

ASJC Scopus subject areas

  • Microbiology
  • Parasitology
  • Virology
  • Immunology
  • Genetics
  • Molecular Biology

Cite this

Role of acetyl-phosphate in activation of the Rrp2-RpoN-RpoS pathway in borrelia burgdorferi. / Xu, Haijun; Caimano, Melissa J.; Lin, Tao; He, Ming; Radolf, Justin D.; Norris, Steven J.; Gheradini, Frank; Wolfe, Alan J.; Yang, X.

In: PLoS Pathogens, Vol. 6, No. 9, e01104, 09.2010.

Research output: Contribution to journalArticle

Xu, H, Caimano, MJ, Lin, T, He, M, Radolf, JD, Norris, SJ, Gheradini, F, Wolfe, AJ & Yang, X 2010, 'Role of acetyl-phosphate in activation of the Rrp2-RpoN-RpoS pathway in borrelia burgdorferi', PLoS Pathogens, vol. 6, no. 9, e01104. https://doi.org/10.1371/journal.ppat.1001104
Xu, Haijun ; Caimano, Melissa J. ; Lin, Tao ; He, Ming ; Radolf, Justin D. ; Norris, Steven J. ; Gheradini, Frank ; Wolfe, Alan J. ; Yang, X. / Role of acetyl-phosphate in activation of the Rrp2-RpoN-RpoS pathway in borrelia burgdorferi. In: PLoS Pathogens. 2010 ; Vol. 6, No. 9.
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abstract = "Borrelia burgdorferi, the Lyme disease spirochete, dramatically alters its transcriptome and proteome as it cycles between the arthropod vector and mammalian host. During this enzootic cycle, a novel regulatory network, the Rrp2-RpoN-RpoS pathway (also known as the σ54-σS sigma factor cascade), plays a central role in modulating the differential expression of more than 10{\%} of all B. burgdorferi genes, including the major virulence genes ospA and ospC. However, the mechanism(s) by which the upstream activator and response regulator Rrp2 is activated remains unclear. Here, we show that none of the histidine kinases present in the B. burgdorferi genome are required for the activation of Rrp2. Instead, we present biochemical and genetic evidence that supports the hypothesis that activation of the Rrp2-RpoN-RpoS pathway occurs via the small, high-energy, phosphoryl-donor acetyl phosphate (acetyl~P), the intermediate of the Ack-Pta (acetate kinase-phosphate acetyltransferase) pathway that converts acetate to acetyl-CoA. Supplementation of the growth medium with acetate induced activation of the Rrp2-RpoN-RpoS pathway in a dose-dependent manner. Conversely, the overexpression of Pta virtually abolished acetate-induced activation of this pathway, suggesting that acetate works through acetyl,P. Overexpression of Pta also greatly inhibited temperature and cell density-induced activation of RpoS and OspC, suggesting that these environmental cues affect the Rrp2-RpoN-RpoS pathway by influencing acetyl~P. Finally, overexpression of Pta partially reduced infectivity of B. burgdorferi in mice. Taken together, these findings suggest that acetyl~P is one of the key activating molecule for the activation of the Rrp2-RpoN-RpoS pathway and support the emerging concept that acetyl~P can serve as a global signal in bacterial pathogenesis.",
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