Rrp2, a σ
 54
 -Dependent Transcriptional Activator of
 Borrelia burgdorferi
 , Activates
 rpoS
 in an Enhancer-Independent Manner

Blevins, Jon S.; Xu, Hai Jun; He, Ming; Norgard, Michael V.; Reitzer, Larry; Yang, Xiaoming

doi:10.1128/jb.01721-08

Cited by 51 publications

(82 citation statements)

References 27 publications

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“…5c), suggesting that the decrease of RpoS in the csrA Bb mutant occurs upstream of the RpoS pathway. The expression of rpoS is directly controlled by the alternative sigma factor RpoN, whose expression requires the activation of the response regulator Rrp2 (9,55). In addition, recent studies showed that BosR, a homolog of the Fur regulatory protein, interfaces with the Rrp2-RpoN-RpoS network (24,38).…”

Section: Resultsmentioning

confidence: 99%

“…Rrp2-RpoNRpoS forms a central regulatory network of B. burgdorferi (11,23,66). RpoN directly controls the expression of RpoS, and Rrp2 is an activator of the RpoN-RpoS pathway (9,66). In addition, recent studies showed that BosR, a homolog of the Fur regulatory protein, interfaces with the Rrp2-RpoN-RpoS network (24,38).…”

Section: Discussionmentioning

confidence: 99%

“…B. burgdorferi cells were cultivated at 34°C/pH 7.6 and harvested at stationary phase (ϳ10 8 cells/ml). Equal amounts of whole-cell lysates (10 to 50 g) were separated on an SDS-PAGE gel and transferred to a polyvinylidene difluoride (PVDF) membrane (Bio-Rad Labo- Upstream of csrA Bb (R) 5Ј-TCTAGATGATTGGTGCCTTTAGGTTAG-3Ј P 3 Downstream of csrA Bb (F) 5Ј-TCTAGATAACCTCTGCATTTTGTC-3Ј P 4 Downstream of csrA Bb (R) 5Ј-CATATGTTCTTTGGAAGAATTTGAGC-3Ј P 5 Kan (F) 5Ј-TCTAGATAATACCCGAGCTTCAAG-3Ј P 6 Kan (R) 5Ј-TCTAGATCAAGTCAGCGTAATGCTCTG-3Ј P 7 Complementation, P flgK (F) 5Ј-GGATCCGCACTACTTAAAAAAGGTGTTGC-3Ј P 8 Complementation, P flgK (R) 5Ј-CATATGTTTTATGAAATTAATTATAAGC-3Ј P 9 Complementation, csrA Bb (F) 5Ј-CATATGCTAGTATTGTCAAGAAAAGC-3Ј P 10 Complementation, csrA Bb (R) 5Ј-GGATCCTTATTTGTCATCGTCGTCC-3Ј P 11 qRT-PCR, rpoS (F) 5Ј-ACCTATCTCCTGCTCAGTATATAA-3Ј P 12 qRT-PCR, rpoS (R) 5Ј-CAAGGGTAATTTCAGGGTTAAAAG-3Ј P 13 qRT-PCR, ospC (F) 5Ј-TGTTACTGATGCTGATGCAA-3Ј P 14 qRT-PCR, ospC (R) 5Ј-AAGCTCTTTAACTGAATTAGC-3Ј P 15 qRT-PCR, dbpA (F) 5Ј-GGACTAACAGGAGCAACA-3Ј P 16 qRT-PCR, dbpA (R) 5Ј-CACCACTACTTCCAGTTTC-3Ј P 17 qRT-PCR, ospA (F) 5Ј-GCAGCCTTGACGAGAAAAACA-3Ј P 18 qRT-PCR, ospA (R) 5Ј-CGCCTTCAAGTACTCCAGATCC-3Ј P 19 qRT-PCR, dsrA (F) 5Ј-AATGAAGTTAGTGGGCGTTACTC-3Ј P 20 qRT-PCR, dsrA (R) 5Ј-TTTTTTTGAATAGGGTCACCAG-3Ј P 21 qRT-PCR, eno (F) 5Ј-AACAGGAATTAACGAGGCTG-3Ј P 22 qRT-PCR, eno (R) 5Ј-AAATTGCATTAGCACCAAGC-3Ј P 23 qRT-PCR, bb0589 (F) 5Ј-GAGTTTTAAAGGCAGCTATTGT-3Ј P 24 qRT-PCR, bb0589 (R) 5Ј-CTTTGCTTCGTAACTCCCTA-3Ј P 25 Co-RT, bb0588 (F) 5Ј-CTGCTTTCAATTCAGCCAAAG-3Ј P 26 Co-RT, bb0589 (R) 5Ј-GCCTATCAAAATAATCGAATCTGC-3Ј P 27 rCsrA Bb (F) 5Ј-CACCATGCTAGTATTGTCAAGAAAAG-3Ј P 28 rCsrA Bb (R) 5Ј-ATTTTCATTCTTGAAATAATG-3Ј P 29 EMSA probe 5Ј-UUUAUUAUAAGGAGUGUGAUUUU-3Ј P 30 EMSA probe (mutated) 5Ј-UUUAUUAUAAAAAGUGUGAUUUU-3Ј…”

Section: Methodsmentioning

confidence: 99%

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Inactivation of bb0184 , Which Encodes Carbon Storage Regulator A, Represses the Infectivity of Borrelia burgdorferi

Sze

2011

Infect Immun

106

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The genome of Borrelia burgdorferi, the Lyme disease spirochete, encodes a homolog (the bb0184 gene product) of the carbon storage regulator A protein (CsrA Bb ); recent studies reported that CsrA Bb is involved in the regulation of several infectivity factors of B. burgdorferi. However, the mechanism involved remains unknown. In this report, a csrA Bb mutant was constructed and complemented in an infectious B31A3 strain. Subsequent animal studies showed that the mutant failed to establish an infection in mice, highlighting that CsrA Bb is required for the infectivity of B. burgdorferi. Western blot analyses revealed that the virulenceassociated factors OspC, DbpB, and DbpA were attenuated in the csrA Bb mutant. The Rrp2-RpoN-RpoS pathway ( 54 -S sigma factor cascade) is a central regulon that governs the expression of ospC, dbpB, and dbpA. Further analyses found that the level of RpoS was significantly decreased in the mutant, while the level of Rrp2 remained unchanged. A recent study reported that the overexpression of BB0589, a phosphate acetyl-transferase (Pta) that converts acetyl-phosphate to acetyl-coenzyme A (CoA), led to the inhibition of RpoS and OspC expression, suggesting that acetyl-phosphate is an activator of Rrp2. Along with this report, we found that CsrA Bb binds to the leader sequence of the bb0589 transcript and that the intracellular level of acetyl-CoA in the csrA Bb mutant was significantly increased compared to that of the wild type, suggesting that more acetyl-phosphate was being converted to acetyl-CoA in the mutant. Collectively, these results suggest that CsrA Bb may influence the infectivity of B. burgdorferi via regulation of acetate metabolism and subsequent activation of the Rrp2-RpoN-RpoS pathway.Borrelia burgdorferi, the causative agent of Lyme borreliosis, has a complex natural enzootic life cycle-transmitting between Ixodes tick vectors and mammals (56, 57). As such, differential gene expression plays an important role in its adaptation to diverse host environments (10, 45). To date, a limited number of regulatory pathways have been identified in B. burgdorferi (13,16,23,34,38,46,66). Among these identified regulatory factors, the Rrp2-RpoN-RpoS pathway is a central regulatory network of B. burgdorferi, which consists of a two-component response regulator, Rrp2, and two alternative sigma factors, RpoN ( 54 ) and RpoS ( S

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Inactivation of bb0184 , Which Encodes Carbon Storage Regulator A, Represses the Infectivity of Borrelia burgdorferi

Sze

2011

Infect Immun

106

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“…Signals contained within the nymphal blood meal trigger a complex series of transcriptional, antigenic, and physiological changes that enable spirochetes colonizing the midgut to disseminate through tick tissues while preparing for growth within the mammalian host (11,15,16,67); the Rrp2/RpoN/RpoS regulatory pathway is central to this transmission-associated program (6,7,11,15,20,52,86,87). The signaling pathways underlying the converse process, whereby spirochetes transit from the mammalian host into naïve I. scapularis ticks, have received comparatively little attention.…”

Section: Discussionmentioning

confidence: 99%

“…Although Hk2 was widely presumed to be the cognate HK for Rrp2, Xu et al (86) recently demonstrated that the high-energy, phosphoryl donor acetyl phosphate (acetylϳP) is capable of phosphorylating Rrp2 in vitro and, more importantly, that Hk2 is not required for activation of Rrp2 in vivo. Once activated, Rrp2ϳP acts as a transcriptional activator for the alternate sigma factor RpoN (6,87), which in turn controls expression of the spirochete's other alternate sigma factor, RpoS (7,20,36,52,70). Genes within the Rrp2/ RpoN/RpoS regulon promote tick-to-mammal transmission (30) and early murine infection (10,20,32,68,80,83).…”

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confidence: 99%

The Hybrid Histidine Kinase Hk1 Is Part of a Two-Component System That Is Essential for Survival of Borrelia burgdorferi in Feeding Ixodes scapularis Ticks

et al. 2011

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Two-component systems (TCS) are principal mechanisms by which bacteria adapt to their surroundings.Borrelia burgdorferi encodes only two TCS. One is comprised of a histidine kinase, Hk2, and the response regulator Rrp2. While the contribution of Hk2 remains unclear, Rrp2 is part of a regulatory pathway involving the spirochete's alternate sigma factors, RpoN and RpoS. Genes within the Rrp2/RpoN/RpoS regulon function to promote tick transmission and early infection. The other TCS consists of a hybrid histidine kinase, Hk1, and the response regulator Rrp1. Hk1 is composed of two periplasmic sensor domains (D1 and D2), followed by conserved cytoplasmic histidine kinase core, REC, and Hpt domains. In addition to its REC domain, Rrp1 contains a GGDEF motif characteristic of diguanylate cyclases. To investigate the role of Hk1 during the enzootic cycle, we inactivated this gene in two virulent backgrounds. Extensive characterization of the resulting mutants revealed a dramatic phenotype whereby Hk1-deficient spirochetes are virulent in mice and able to migrate out of the bite site during feeding but are killed within the midgut following acquisition. We hypothesize that the phosphorelay between Hk1 and Rrp1 is initiated by the binding of feeding-specific ligand(s) to Hk1 sensor domain D1 and/or D2. Once activated, Rrp1 directs the synthesis of cyclic dimeric GMP (c-di-GMP), which, in turn, modulates the expression and/or activity of gene products required for survival within feeding ticks. In contrast to the Rrp2/RpoN/RpoS pathway, which is active only within feeding nymphs, the Hk1/Rrp1 TCS is essential for survival during both larval and nymphal blood meals.

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Insights into the Biology of Borrelia burgdorferi Gained Through the Application of Molecular Genetics

Groshong

Blevins

2014

Advances in Applied Microbiology

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Rrp2, a σ ⁵⁴ -Dependent Transcriptional Activator of Borrelia burgdorferi , Activates rpoS in an Enhancer-Independent Manner

Cited by 51 publications

References 27 publications

Inactivation of bb0184 , Which Encodes Carbon Storage Regulator A, Represses the Infectivity of Borrelia burgdorferi

Inactivation of bb0184 , Which Encodes Carbon Storage Regulator A, Represses the Infectivity of Borrelia burgdorferi

The Hybrid Histidine Kinase Hk1 Is Part of a Two-Component System That Is Essential for Survival of Borrelia burgdorferi in Feeding Ixodes scapularis Ticks

Insights into the Biology of Borrelia burgdorferi Gained Through the Application of Molecular Genetics

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Rrp2, a σ 54 -Dependent Transcriptional Activator of Borrelia burgdorferi , Activates rpoS in an Enhancer-Independent Manner

Cited by 51 publications

References 27 publications

Inactivation of bb0184 , Which Encodes Carbon Storage Regulator A, Represses the Infectivity of Borrelia burgdorferi

Inactivation of bb0184 , Which Encodes Carbon Storage Regulator A, Represses the Infectivity of Borrelia burgdorferi

The Hybrid Histidine Kinase Hk1 Is Part of a Two-Component System That Is Essential for Survival of Borrelia burgdorferi in Feeding Ixodes scapularis Ticks

Insights into the Biology of Borrelia burgdorferi Gained Through the Application of Molecular Genetics

Contact Info

Product

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Rrp2, a σ ⁵⁴ -Dependent Transcriptional Activator of Borrelia burgdorferi , Activates rpoS in an Enhancer-Independent Manner