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Host-specific functional compartmentalization within the oligopeptide transporter during the Borrelia burgdorferi enzootic cycle.

ABSTRACT: Borrelia burgdorferi must acquire all of its amino acids (AAs) from its arthropod vector and vertebrate host. Previously, we determined that peptide uptake via the oligopeptide (Opp) ABC transporter is essential for spirochete viability in vitro and during infection. Our prior study also suggested that B. burgdorferi employs temporal regulation in concert with structural variation of oligopeptide-binding proteins (OppAs) to meet its AA requirements in each biological niche. Herein, we evaluated the contributions to the B. burgdorferi enzootic cycle of three of the spirochete's five OppAs (OppA1, OppA2, and OppA5). An oppA1 transposon (tn) mutant lysed in the hyperosmolar environment of the feeding tick, suggesting that OppA1 imports amino acids required for osmoprotection. The oppA2tn mutant displayed a profound defect in hematogenous dissemination in mice, yet persisted within skin while inducing only a minimal antibody response. These results, along with slightly decreased growth of the oppA2tn mutant within DMCs, suggest that OppA2 serves a minor nutritive role, while its dissemination defect points to an as yet uncharacterized signaling function. Previously, we identified a role for OppA5 in spirochete persistence within the mammalian host. We now show that the oppA5tn mutant displayed no defect during the tick phase of the cycle and could be tick-transmitted to naïve mice. Instead of working in tandem, however, OppA2 and OppA5 appear to function in a hierarchical manner; the ability of OppA5 to promote persistence relies upon the ability of OppA2 to facilitate dissemination. Structural homology models demonstrated variations within the binding pockets of OppA1, 2, and 5 indicative of different peptide repertoires. Rather than being redundant, B. burgdorferi's multiplicity of Opp binding proteins enables host-specific functional compartmentalization during the spirochete lifecycle.

SUBMITTER: Groshong AM

PROVIDER: S-EPMC7822543 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature

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Host-specific functional compartmentalization within the oligopeptide transporter during the Borrelia burgdorferi enzootic cycle.

Groshong Ashley M AM McLain Melissa A MA Radolf Justin D JD

PLoS pathogens 20210111 1

Borrelia burgdorferi must acquire all of its amino acids (AAs) from its arthropod vector and vertebrate host. Previously, we determined that peptide uptake via the oligopeptide (Opp) ABC transporter is essential for spirochete viability in vitro and during infection. Our prior study also suggested that B. burgdorferi employs temporal regulation in concert with structural variation of oligopeptide-binding proteins (OppAs) to meet its AA requirements in each biological niche. Herein, we evaluated ...[more]

PMID: 33428666

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Project description:Borrelia burgdorferi is an extreme amino acid (AA) auxotroph whose genome encodes few free AA transporters and an elaborate oligopeptide transport system (B. burgdorferi Opp [BbOpp]). BbOpp consists of five oligopeptide-binding proteins (OBPs), two heterodimeric permeases, and a heterodimeric nucleotide-binding domain (NBD). Homology modeling based on the crystal structure of liganded BbOppA4 revealed that each OBP likely binds a distinct range of peptides. Transcriptional analyses demonstrated that the OBPs are differentially and independently regulated whereas the permeases and NBDs are constitutively expressed. A conditional NBD mutant failed to divide in the absence of inducer and replicated in an IPTG (isopropyl-?-d-thiogalactopyranoside) concentration-dependent manner. NBD mutants grown without IPTG exhibited an elongated morphotype lacking division septa, often with flattening at the cell center due to the absence of flagellar filaments. Following cultivation in dialysis membrane chambers, NBD mutants recovered from rats not receiving IPTG also displayed an elongated morphotype. The NBD mutant was avirulent by needle inoculation, but infectivity was partially restored by oral administration of IPTG to infected mice. We conclude that peptides are a major source of AAs for B. burgdorferi both in vitro and in vivo and that peptide uptake is essential for regulation of morphogenesis, cell division, and virulence.IMPORTANCEBorrelia burgdorferi, the causative agent of Lyme disease, is an extreme amino acid (AA) auxotroph with a limited repertoire of annotated single-AA transporters. A major issue is how the spirochete meets its AA requirements as it transits between its arthropod vector and mammalian reservoir. While previous studies have confirmed that the B. burgdorferi oligopeptide transport (opp) system is capable of importing peptides, the importance of the system for viability and pathogenesis has not been established. Here, we evaluated the opp system structurally and transcriptionally to elucidate its ability to import a wide range of peptides during the spirochete's enzootic cycle. Additionally, using a novel mutagenesis strategy to abrogate opp transporter function, we demonstrated that peptide uptake is essential for bacterial viability, morphogenesis, and infectivity. Our studies revealed a novel link between borrelial physiology and virulence and suggest that peptide uptake serves an intracellular signaling function regulating morphogenesis and division.

Project description:In the northeastern U.S., Borrelia burgdorferi sensu stricto, the agent of Lyme disease, is maintained between vertebrate hosts and subadult deer ticks (the northern clade of Ixodes scapularis, formerly known as Ixodes dammini). Theoretical arguments suggest that the force of transmission would be greatest when infected nymphal ticks focus their bites on the same host as the uninfected larvae. Stage-specific differences in host utilization would reduce the force of transmission, but to date such differences remain understudied. We determined the host utilization differences of larval and nymphal deer ticks using bloodmeal analysis of host-seeking nymphs and adults collected from 5 field sites in New England. Matched cohorts of ticks (nymphs=506, adults=451), i.e. ticks that had fed during the same summer season, were used to control for yearly host population variations. Infection status of all ticks was determined by real time PCR. Nymphal deer ticks were more likely to have fed on birds and sciurids (13% vs 3%, and 41% vs 9%, respectively p<0.001) and larvae were more likely to have fed on shrews (26% vs 3%, p<0.001). Similarly, ticks that had fed on a mouse or a shrew as larvae were likely to become infected (OR= 3.195, 95% CI [1.9, 5.1] and OR=2.5[1.6,3.8] respectively), and they were positively associated with infection prevalence at our sites. However, very few nymphs fed on shrews, and they were not associated with infection, raising the question of how uninfected shrews acquire infection each year. Sciurids did not appear to contribute to the enzootic cycle at our sites, which may be due to the low numbers of larvae that fed on them. Sciurid-fed ticks of either stage were not associated with infection. Both stages of ticks were less likely to be infected if they had fed on deer (OR=0.08 [0.02.0.3] and OR=0.4 [0.2,0.7] tested as nymphs and adults, respectively) and thus deer likely served to reduce the force of transmission at our sites. Site-specific analysis of differential host utilization by subadult deer ticks may contribute to appropriate targeting of interventions and thereby promote reducing risk of acquiring Lyme disease and the other deer tick-transmitted infections.

Dataset Information

Host-specific functional compartmentalization within the oligopeptide transporter during the Borrelia burgdorferi enzootic cycle.

Publications

Host-specific functional compartmentalization within the oligopeptide transporter during the Borrelia burgdorferi enzootic cycle.

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