Quantitative analyses of interactions between SpoVG and RNA/DNA

Saylor, Timothy C.; Savage, Christina R.; Krusenstjerna, Andrew C.; Jusufovic, Nerina; Zückert, Wolfram R.; Brissette, Catherine A.; Motaleb, M. A.; Schlax, Paula J.; Stevenson, Brian

doi:10.1016/j.bbrc.2023.02.044

Cited by 3 publications

(8 citation statements)

References 15 publications

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“…BadR, the B orrelia Host Ad aption R egulator, has recently been identified as a repressor of glpFKD , while PlzA has previously been shown to exert both positive and negative effects on glpFKD , at high and low levels of c-di-GMP respectively (Zhang et al ., 2018; Zhang et al ., 2022). Although SpoVG can bind the DNA and RNA of glpFKD , the transcriptional consequences of these interactions are unknown (Savage et al ., 2018; Saylor et al ., 2023). How PlzA binding affects glpFKD expression, and how these proteins work in tandem to regulate glycerol catabolism, is currently being investigated.…”

Section: Discussionmentioning

confidence: 99%

“…PlzA bound to a sequence adjacent to the promoter and extending 35 bp into the 5’ UTR of glpF . This site has been shown to positively affect glpFKD transcript levels and is a site where two other borrelial regulators, SpoVG and BadR, also bind (Grove et al ., 2017; Savage et al ., 2018; Zhang et al ., 2022; Saylor et al ., 2023). BadR, the B orrelia Host Ad aption R egulator, has recently been identified as a repressor of glpFKD , while PlzA has previously been shown to exert both positive and negative effects on glpFKD , at high and low levels of c-di-GMP respectively (Zhang et al ., 2018; Zhang et al ., 2022).…”

Section: Discussionmentioning

confidence: 99%

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Borrelia burgdorferiPlzA is a cyclic-di-GMP dependent DNA and RNA binding protein

Jusufovic

Savage

Saylor

et al. 2023

Preprint

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The PilZ domain-containing protein, PlzA, is the only cyclic di-GMP binding protein encoded by all Lyme disease spirochetes. PlzA has been implicated in the regulation of many borrelial processes, but the functional mechanism of PlzA was not previously known. We report that PlzA can bind DNA and RNA within the promoter and 5' UTR of the glycerol metabolism operon, glpFKD, and that nucleic acid binding requires c-di-GMP. In the presence of c-di-GMP, PlzA formed multimeric complexes with nucleic acids. PlzA contains two PilZ domains. Dissection of the domains demonstrated that the separated N-terminal domain bound nucleic acids in a c-di-GMP-independent manner. The C-terminal domain, which includes the c-di-GMP binding motif, did not bind nucleic acids under any tested conditions. Structural modeling suggests that c-di-GMP binding to the C-terminal domain stabilizes interactions between the two domains, facilitating nucleic acid binding through residues in the N-terminal domain.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Borrelia burgdorferiPlzA is a cyclic-di-GMP dependent DNA and RNA binding protein

Jusufovic

Savage

Saylor

et al. 2023

Preprint

Self Cite

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show abstract

“…PlzA bound to a sequence adjacent to the promoter and extending 35 bp into the 5′ UTR of glpF. This site has been shown to positively affect glpFKD transcript levels and is a site where two other borrelial regulators, SpoVG and BadR, also bind (Grove et al, 2017;Savage et al, 2018;Saylor et al, 2023;Zhang et al, 2024). BadR, the Borrelia Host Adaption Regulator, has recently been identified as a repressor of glpFKD, while PlzA has previously been shown to exert both positive and negative effects on glpFKD, at high and low levels of c-di-GMP, respectively (Zhang et al, 2018(Zhang et al, , 2024.…”

Section: Discussionmentioning

confidence: 99%

“…to bind the same site within glpFKD, SpoVG and BadR (Savage et al, 2018;Saylor et al, 2023;Zhang et al, 2024). Competition between these proteins could also impact the ChIP data.…”

Section: Discussionmentioning

confidence: 99%

“…Overall, our results suggest that the glp-FKD(−7/+35) sequence may be a non-consensus or low-affinity DNA binding site of PlzA within the glpFKD operon. This 42 bp sequence was chosen as it is crucial for glpFKD regulation and is known to be bound by other proteins (Grove et al, 2017;Savage et al, 2018;Saylor et al, 2023;Zhang et al, 2024). We hypothesize that there could be higher affinity binding sites upstream of this operon and within the B. burgdorferi genome.…”

Section: Despite Introducing Various Mutations Into the Glpfkd(−7/+35)mentioning

confidence: 99%

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Borrelia burgdorferiPlzA is a cyclic‐di‐GMP dependent DNA and RNA binding protein

Jusufovic,

Krusenstjerna,

Savage

et al. 2024

Molecular Microbiology

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The PilZ domain‐containing protein, PlzA, is the only known cyclic di‐GMP binding protein encoded by all Lyme disease spirochetes. PlzA has been implicated in the regulation of many borrelial processes, but the effector mechanism of PlzA was not previously known. Here, we report that PlzA can bind DNA and RNA and that nucleic acid binding requires c‐di‐GMP, with the affinity of PlzA for nucleic acids increasing as concentrations of c‐di‐GMP were increased. A mutant PlzA that is incapable of binding c‐di‐GMP did not bind to any tested nucleic acids. We also determined that PlzA interacts predominantly with the major groove of DNA and that sequence length and G–C content play a role in DNA binding affinity. PlzA is a dual‐domain protein with a PilZ‐like N‐terminal domain linked to a canonical C‐terminal PilZ domain. Dissection of the domains demonstrated that the separated N‐terminal domain bound nucleic acids independently of c‐di‐GMP. The C‐terminal domain, which includes the c‐di‐GMP binding motifs, did not bind nucleic acids under any tested conditions. Our data are supported by computational docking, which predicts that c‐di‐GMP binding at the C‐terminal domain stabilizes the overall protein structure and facilitates PlzA‐DNA interactions via residues in the N‐terminal domain. Based on our data, we propose that levels of c‐di‐GMP during the various stages of the enzootic life cycle direct PlzA binding to regulatory targets.

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The Lyme disease spirochete, Borrelia burgdorferi, as a model vector-borne pathogen: insights on regulation of gene and protein expression

Stevenson

2023

Current Opinion in Microbiology

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Quantitative analyses of interactions between SpoVG and RNA/DNA

Cited by 3 publications

References 15 publications

Borrelia burgdorferiPlzA is a cyclic-di-GMP dependent DNA and RNA binding protein

Borrelia burgdorferiPlzA is a cyclic-di-GMP dependent DNA and RNA binding protein

Borrelia burgdorferiPlzA is a cyclic‐di‐GMP dependent DNA and RNA binding protein

The Lyme disease spirochete, Borrelia burgdorferi, as a model vector-borne pathogen: insights on regulation of gene and protein expression

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