HrpA, a DEAH-Box RNA Helicase, Is Involved in Global Gene Regulation in the Lyme Disease Spirochete

Aydan Salman-Dilgimen,Pierre-Olivier Hardy,Ashley R Dresser,George Chaconas,Ben Adler

doi:10.1371/journal.pone.0022168

Abstract

Spirochetes causing Lyme borreliosis are obligate parasites that can only be found in a tick vector or a vertebrate host. The ability to survive in these two disparate environments requires up and downregulation of specific genes by regulatory circuits that remain largely obscure. In this work on the Lyme spirochete, B. burgdorferi, we show that a disruption of the hrpA gene, which encodes a putative RNA helicase, results in a complete loss in the ability of the spirochetes to infect mice by needle inoculation. Studies of protein expression in culture by 2D gels revealed a change in the expression of 33 proteins in hrpA clones relative to the wild-type parent. Quantitative characterization of protein expression by iTRAQ analysis revealed a total of 187 differentially regulated proteins in an hrpA background: 90 downregulated and 97 upregulated. Forty-two of the 90 downregulated and 65 of the 97 upregulated proteins are not regulated under any conditions by the previously reported regulators in B. burgdorferi (bosR, rrp2, rpoN, rpoS or rrp1). Downregulated and upregulated proteins also fell into distinct functional categories. We conclude that HrpA is part of a new and distinct global regulatory pathway in B. burgdorferi gene expression. Because an HrpA orthologue is present in many bacteria, its participation in global regulation in B. burgdorferi may have relevance in other bacterial species where its function remains obscure. We believe this to be the first report of a role for an RNA helicase in a global regulatory pathway in bacteria. This finding is particularly timely with the recent growth of the field of RNA regulation of gene expression and the ability of RNA helicases to modulate RNA structure and function.

Highlights

Lyme borreliosis is common in the northern hemisphere and is the most frequent tick-borne disease in North America and Europe [1,2]
The causative agents, Borrelia burgdorferi and related species, are obligate parasites that survive through a complex enzootic cycle involving a tick vector and a vertebrate host
RNA regulation has been recently reported in B. burgdorferi; the RNA regulator DsrA has been shown to regulate the expression of rpoS and ospC [28] and the RNA chaperone Hfq appears to be involved in regulating the expression of pathogenicity factors [29]

Summary

Introduction

Lyme borreliosis is common in the northern hemisphere and is the most frequent tick-borne disease in North America and Europe [1,2]. About 150 genes appear to be differentially regulated in B. burgdorferi, depending upon environmental conditions and tick or host factors required for survival in these very different settings [6,7,8,9,10, 11,12,13,14,15,16]. Several different regulatory molecules and pathways that play a role in differential gene expression have been identified, including the BosR regulator [5,14,15,16,17,18], alternative s factors RpoS and RpoN [7,12,19,20] and the RpoN activator Rrp2 [21,22,23]. RNA regulation has been recently reported in B. burgdorferi; the RNA regulator DsrA has been shown to regulate the expression of rpoS and ospC [28] and the RNA chaperone Hfq appears to be involved in regulating the expression of pathogenicity factors [29]

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Conclusion