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Genome-wide screen identifies novel genes required for Borrelia burgdorferi survival in its Ixodes tick vector.


ABSTRACT: Borrelia burgdorferi, the causative agent of Lyme disease in humans, is maintained in a complex biphasic life cycle, which alternates between tick and vertebrate hosts. To successfully survive and complete its enzootic cycle, B. burgdorferi adapts to diverse hosts by regulating genes required for survival in specific environments. Here we describe the first ever use of transposon insertion sequencing (Tn-seq) to identify genes required for B. burgdorferi survival in its tick host. We found that insertions into 46 genes resulted in a complete loss of recovery of mutants from larval Ixodes ticks. Insertions in an additional 56 genes resulted in a >90% decrease in fitness. The screen identified both previously known and new genes important for larval tick survival. Almost half of the genes required for survival in the tick encode proteins of unknown function, while a significant portion (over 20%) encode membrane-associated proteins or lipoproteins. We validated the results of the screen for five Tn mutants by performing individual competition assays using mutant and complemented strains. To better understand the role of one of these genes in tick survival, we conducted mechanistic studies of bb0017, a gene previously shown to be required for resistance against oxidative stress. In this study we show that BB0017 affects the regulation of key borrelial virulence determinants. The application of Tn-seq to in vivo screening of B. burgdorferi in its natural vector is a powerful tool that can be used to address many different aspects of the host pathogen interaction.

SUBMITTER: Phelan JP 

PROVIDER: S-EPMC6516651 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature

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Genome-wide screen identifies novel genes required for Borrelia burgdorferi survival in its Ixodes tick vector.

Phelan James P JP   Kern Aurelie A   Ramsey Meghan E ME   Lundt Maureen E ME   Sharma Bijaya B   Lin Tao T   Gao Lihui L   Norris Steven J SJ   Hyde Jenny A JA   Skare Jon T JT   Hu Linden T LT  

PLoS pathogens 20190514 5


Borrelia burgdorferi, the causative agent of Lyme disease in humans, is maintained in a complex biphasic life cycle, which alternates between tick and vertebrate hosts. To successfully survive and complete its enzootic cycle, B. burgdorferi adapts to diverse hosts by regulating genes required for survival in specific environments. Here we describe the first ever use of transposon insertion sequencing (Tn-seq) to identify genes required for B. burgdorferi survival in its tick host. We found that  ...[more]

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