Development of Genetic System to Inactivate a Borrelia turicatae Surface Protein Selectively Produced within the Salivary Glands of the Arthropod Vector

Abstract

Background

Borrelia turicatae, an agent of tick-borne relapsing fever, is an example of a pathogen that can adapt to disparate conditions found when colonizing the mammalian host and arthropod vector. However, little is known about the genetic factors necessary during the tick-mammalian infectious cycle, therefore we developed a genetic system to transform this species of spirochete. We also identified a plasmid gene that was up-regulated in vitro when B. turicatae was grown in conditions mimicking the tick environment. This 40 kilodalton protein was predicted to be surface localized and designated the Borrelia repeat protein A ( brpA) due to the redundancy of the amino acid motif Gln-Gly-Asn-Val-Glu.

Methodology/Principal Findings

Quantitative reverse-transcriptase polymerase chain reaction using RNA from B. turicatae infected ticks and mice indicated differential regulation of brpA during the tick-mammalian infectious cycle. The surface localization was determined, and production of the protein within the salivary glands of the tick was demonstrated. We then applied a novel genetic system for B. turicatae to inactivate brpA and examined the role of the gene product for vector colonization and the ability to establish murine infection.

Conclusions/Significance

These results demonstrate the complexity of protein production in a population of spirochetes within the tick. Additionally, the development of a genetic system is important for future studies to evaluate the requirement of specific B. turicatae genes for vector colonization and transmission.

Author Summary

Relapsing fever spirochetes are a global yet neglected pathogen causing recurrent febrile episodes, nausea, vomiting, and pregnancy complications including miscarriage. Most species of tick-borne relapsing fever spirochetes are maintained in enzootic cycles, and given an approximately 20 year life span, the arthropod vector for Borrelia turicatae represents a reservoir for the pathogens. While B. turicatae has adapted mechanisms to efficiently colonize and survive within the vector, the genes necessary during the tick-mammalian infectious cycle are unknown. We have identified a gene that was designated the Borrelia repeat protein A ( brpA). brpA was up-regulated in a portion of the spirochetes colonizing Ornithodoros turicata, the vector for B. turicatae. Developing a system to delete the gene in B. turicatae enabled the evaluation of the necessity of brpA. With the genetic system established for B. turicatae, a better understanding of the genetic constituents required during the tick-mammalian infectious cycle may be obtained.