Insight into the dual functions of bacterial enhancer-binding protein Rrp2 of Borrelia burgdorferi

Yanping Yin, Youyun Yang, Xuwu Xiang, Qian Wang, Zhang Nv Yang, Jon Blevins, Yongliang Lou, X. Frank Yang

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

It is well established that the RpoN-RpoS sigma factor (∂54-∂S) cascade plays an essential role in differential gene expression during the enzootic cycle of Borrelia burgdorferi, the causative agent of Lyme disease. The RpoN-RpoS pathway is activated by the response regulator/∂54-dependent activator (also called bacterial enhancer-binding protein [bEBP]) Rrp2. One unique feature of Rrp2 is that this activator is essential for cell replication, whereas RpoN-RpoS is dispensable for bacterial growth. How Rrp2 controls cell replication, a function that is independent of RpoN-RpoS, remains to be elucidated. In this study, by generating a series of conditional rrp2 mutant strains, we demonstrated that the N-terminal receiver domain of Rrp2 is required for spirochetal growth. Furthermore, a D52A point mutation at the phosphorylation site within the N terminus of Rrp2 abolished cell replication. Mutation of the ATPase motif within the central domain of Rrp2 did not affect spirochetal replication, indicating that phosphorylation-dependent ATPase activity of Rrp2 for ∂54 activation is not required for cell growth. However, deletion of the C-terminal domain or a 16-amino-acid truncation of the helix-turn-helix (HTH) DNA-binding motif within the C-terminal domain of Rrp2 abolished spirochetal replication. It was shown that constitutive expression of rpoS is deleterious to borrelial growth. We showed that the essential nature of Rrp2 is not due to an effect on rpoS. These data suggest that phosphorylationdependent oligomerization and DNA binding of Rrp2 likely function as a repressor, independently of the activation of ∂54, controlling an essential step of cell replication in B. burgdorferi.

Original languageEnglish (US)
Pages (from-to)1543-1552
Number of pages10
JournalJournal of bacteriology
Volume198
Issue number10
DOIs
StatePublished - May 1 2016

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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