Biology subjects

Hyde, J. A.

Publications and source records attributed to Hyde, J. A..

4 recordsLinked to original sources

bioRxiv2026

Borrelia burgdorferi bb0164 encodes a Na+/Ca2+ antiporter homolog with a novel role in Mn2+ homeostasis and infectivity

Borrelia burgdorferi, the Lyme disease causative agent, relies on trace metals for motility, growth, and virulence in the absence of metal transport homologues encoded in the genome. Previous studies characterized borrelial metal transporter (bmtA) as a manganese (Mn) transporter and speculated that it is the sole Mn transporter. An oxidative stress transposon library screen identified bb0164, annotated as a calcium/sodium antiporter, as having a putative metal binding domain. The transposon mutant lost the ability to internalize Mn suggesting B. burgdorferi is using a non-canonical protein for metal transport. In this study, a bb0164 deletion and complement were generated in B. burgdorferi 5A4-NP1 to evaluate for trace metal transport, virulence regulation, resistance to oxidative stress, and infectivity. Our data demonstrated that the loss of bb0164 resulted in a significant reduction in internalized Mn, increased sensitivity to oxidative stress, dysregulation of the BosR-RpoS virulence pathway, and a loss of infectivity in mice. The loss of bb0164 resulted in elevated rpoS, ospC, and dbpA expression and production, while bosR was not altered transcriptionally or post-transcriptionally. B. burgdorferi grown in chelated complete BSK-II media showed a similar sensitivity to oxidative stress and virulence dysregulation as the bb0164 mutant. These phenotypes were rescued by exogenous Mn and Zn without influencing the expression levels of bb0164 or bmtA. AlphaFold models of BB0164 were structurally divergent from the canonical bacterial Mn transporter, Bacillus subtilis MntH and B. burgdorferi BmtA, but shared high similarity with a calcium/cation antiporter superfamily member. Together, this study characterized BB0164 as a second non-canonical Mn transporter in B. burgdorferi that is essential for mammalian pathogenesis and likely supports metal homeostasis along with bmtA. More broadly, B. burgdorferi uses unique and uncharacterized mechanisms for metal homeostasis that supports physiology and pathogenesis of the spirochete during mammalian infection. Author SummaryLyme disease, caused by Borrelia burgdorferi, is the most common vector-borne illness in the United States and can result in a chronic inflammatory disease. B. burgdorferi acquires most of the necessary nutrients, including trace metals, from the host due to its limited metabolic capacity. B. burgdorferi has evolved a manganese-centric metabolism in place of the iron primarily used by other bacteria. Little is understood about manganese homeostasis in B. burgdorferi with a single transporter, BmtA, characterized to date. Here, we describe a second manganese transporter, encoded by bb0164, that is essential for infection, protects against oxidative stress, and alters genetic regulation. We found that BB0164, an annotated ion antiporter, does not structurally align with conserved manganese transporters from other bacteria. Interestingly, our work suggests bb0164 and bmtA are not subject to transcriptional regulation dependent on temperature or metal availability, differing from other bacterial manganese transporters. These findings indicate B. burgdorferi is uniquely using an antiporter protein for metal transport to support metal homeostasis, which further demonstrates the importance of manganese in borrelial pathogenesis.

microbiology↗

bioRxiv2025

Identification of proteins exhibiting in vitro RNA chaperone activity through gradient profiling in the Lyme disease spirochete, Borrelia burgdorferi

RNA-binding proteins (RBPs) play key roles in regulating gene expression in bacteria. However, relatively few RBPs have been discovered and characterized in Borrelia (Borreliella) burgdorferi, the causative agent of Lyme disease. We utilized gradient profiling to identify putative RBPs that co-sediment with small RNAs (sRNAs) and nascent mRNAs. We employed in vitro and in vivo assays to characterize the RNA chaperone activities of several proteins we identified in the gradient profiling. The previously hypothetical proteins BB0749, BB0713 and BB0796, as well as the chemotaxis-related protein CheY2 and the flagella-associated protein FlgV displayed RNA annealing and/or strand displacement activity. Moreover, in vivo Co-IP assays demonstrated BB0749 binds RNA in B. burgdorferi.

molecular biology↗

bioRxiv2022

Borrelia burgdorferi engages mammalian type I interferon responses via the cGAS-STING pathway

Borrelia burgdorferi, the etiologic agent of Lyme disease, is a spirochete that modulates numerous host pathways to cause a chronic, multi-system inflammatory disease in humans. B. burgdorferi infection can lead to Lyme carditis, neurologic complications, and arthritis, due to the ability of specific borrelial strains to disseminate, invade, and drive inflammation. B. burgdorferi elicits type I interferon (IFN-I) responses in mammalian cells and tissues that are associated with the development of severe arthritis or other Lyme-related complications. However, the innate immune sensors and signaling pathways controlling IFN-I induction remain unclear. In this study, we examined whether intracellular nucleic acid sensing is required for the induction of IFN-I to B. burgdorferi. Using fluorescence microscopy, we show that B. burgdorferi associates with mouse and human cells in culture and we document that internalized spirochetes co-localize with the pattern recognition receptor cyclic GMP-AMP synthase (cGAS). Moreover, we report that IFN-I responses in mouse macrophages and murine embryonic fibroblasts are significantly attenuated in the absence cGAS or its adaptor Stimulator of Interferon Genes (STING), which function to sense and respond to intracellular DNA. Longitudinal in vivo tracking of bioluminescent B. burgdorferi revealed similar dissemination kinetics and borrelial load in C57BL/6J wild-type, cGAS-deficient, or STING-deficient mice. However, infection-associated tibiotarsal joint pathology and inflammation were modestly reduced in cGAS-deficient compared to wild-type mice. Collectively, these results indicate that the cGAS-STING pathway is a critical mediator of mammalian IFN-I signaling and innate immune responses to B. burgdorferi. KEY POINTSO_LIB. burgdorferi triggers type I interferon responses in macrophages and fibroblasts C_LIO_LICoiled spirochetes are observed in the cytosol and co-localize with cGAS C_LIO_LIcGAS and STING mediate B. burgdorferi-induced type I interferon responses C_LI

immunology↗

bioRxiv2021

The Borrelia burgdorferi adenylyl cyclase, CyaB, is important for virulence factor production and mammalian infection

Borrelia burgdorferi, the causative agent of Lyme disease, traverses through vastly distinct environments between the tick vector and the multiple phases of the mammalian infection that requires genetic adaptation for the progression of pathogenesis. Borrelial gene expression is highly responsive to changes in specific environmental signals that initiate the RpoS regulon for mammalian adaptation, but the mechanism(s) for direct detection of environmental cues has yet to be identified. Secondary messenger cyclic adenosine monophosphate (cAMP) produced by adenylate cyclase is responsive to environmental signals, such as carbon source and pH, in many bacterial pathogens to promote virulence by altering gene regulation. B. burgdorferi encodes a single non-toxin class IV adenylate cyclase (bb0723, cyaB). This study investigates cyaB expression along with its influence on borrelial virulence regulation and mammalian infectivity. Expression of cyaB was specifically induced with co-incubation of mammalian host cells that was not observed with cultivated tick cells suggesting that cyaB expression is influenced by cellular factor(s) unique to mammalian cell lines. The 3 end of cyaB also encodes a small RNA, SR0623, in the same orientation that overlaps with bb0722. The differential processing of cyaB and SR0623 transcripts may alter the ability to influence function in the form of virulence determinant regulation and infectivity. Two independent cyaB deletion B31 strains were generated in 5A4-NP1 and ML23 backgrounds and complemented with the cyaB ORF alone that truncates SR0623, cyaB with intact SR0623, or cyaB with a mutagenized full length SR0623 to evaluate the influence on transcriptional and post-transcriptional regulation of borrelial virulence factors and infectivity. In the absence of cyaB, expression and production of ospC was significantly reduced, while the protein levels for BosR and DbpA were substantially lower than parental strains. Infectivity studies with both independent cyaB mutants demonstrated an attenuated phenotype with reduced colonization of tissues during early disseminated infection. This work suggests that B. burgdorferi utilizes cyaB and potentially cAMP as a regulatory pathway to modulate borrelial gene expression and protein production to promote borrelial virulence and dissemination in the mammalian host.

microbiology↗