Overexpression of CsrA (BB0184) Alters the Morphology and Antigen Profiles of<i>Borrelia burgdorferi</i>

Sanjuán, Eva; Esteve-Gassent, Maria D.; Maruskova, Mahulena; Seshu, J.

doi:10.1128/iai.00673-09

Cited by 41 publications

(97 citation statements)

References 89 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2D and E, wt, mt, ct, lane 2). We found that the levels of OspA, P66 (a surface-exposed integrin binding porin), and NapA were similar in all three strains and did not exhibit any difference upon propagation of these strains under our experimental conditions (53,70). Contrary to a recent observation (38), we did not observe a change in the levels of NapA under growth conditions that resulted in reduced levels of BosR either in the mutant or in the control strains (Fig.…”

Section: Resultscontrasting

confidence: 55%

“…Whole-cell lysates were prepared and separated on SDS-12.5% PAGE gels as described previously (72). The separated proteins were either visualized by Coomassie brilliant blue staining or transferred to polyvinylidene difluoride (PVDF) membranes and subjected to immunoblot analysis as described previously (26,41,50,70). The membranes were probed with ␣-OspA monoclonal antibodies, ␣-BBA34, ␣-BBK32, ␣-DbpA, ␣-SodA, ␣-P66, ␣-BBA64, ␣-BosR, ␣-CsrA Bb , ␣-RpoS, and ␣-NapA antibodies.…”

Section: Methodsmentioning

confidence: 99%

“…The accumulation of this metabolite is partly dependent on the repression of levels of an enzyme, phosphate acetyltransferase (Pta), which converts acetyl phosphate to acetyl-coenzyme A (CoA), leading to conditions that could potentially dampen the activation of the Rrp2-RpoNRpoS pathway. The repression of Pta has been shown to be mediated by CsrA Bb (carbon storage regulator A of B. burgdorferi), which we and others recently demonstrated plays a crucial role in affecting differential gene expression in B. burgdorferi that facilitates adaptation to vertebrate host-specific conditions (41,70,82). Notably, deletion of csrA Bb leads to a reduction in the levels of expression of several members of the rpoS regulon.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Oligopeptide Permease A5 Modulates Vertebrate Host-Specific Adaptation of Borrelia burgdorferi

et al. 2011

Self Cite

View full text Add to dashboard Cite

Borrelia burgdorferi, the agent of Lyme disease, undergoes rapid adaptive gene expression in response to signals unique to its arthropod vector or vertebrate hosts. Among the upregulated genes under vertebrate host conditions is one of the five annotated homologs of oligopeptide permease A (OppA5, BBA34). A mutant lacking oppA5 was constructed in an lp25-deficient isolate of B. burgdorferi strain B31, and the minimal regions of infectivity were restored via a shuttle vector pBBE22 with or without an intact copy of bba34. Immunoblot analysis of the bba34 mutant revealed a reduction in the levels of RpoS, BosR, and CsrA Bb with a concomitant reduction in the levels of OspC, DbpA, BBK32, and BBA64. There were no changes in the levels of OspA, NapA, P66, and three other OppA orthologs. Quantitative transcriptional analysis correlated with the changes in the protein levels. However, the bba34 mutant displayed comparable infectivities in the C3H/HeN mice and the wild-type strain, despite the reduction in several pathogenesis-related proteins. Supplementation of the growth medium with increased levels of select components, notably sodium acetate and sodium bicarbonate, restored the levels of several proteins in the bba34 mutant to wild-type levels. We speculate that the transport of acetate appears to contribute to the accumulation of key metabolites, like acetyl phosphate, that facilitate the adaptation of B. burgdorferi to the vertebrate host by the activation of the Rrp2-RpoN-RpoS pathway. These studies underscore the importance of solute transport to host-specific adaptation of B. burgdorferi.

show abstract

Section: Resultscontrasting

confidence: 55%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Oligopeptide Permease A5 Modulates Vertebrate Host-Specific Adaptation of Borrelia burgdorferi

et al. 2011

Self Cite

View full text Add to dashboard Cite

show abstract

“…b Normal periplasmic flagella were tilted toward the cell body. 43,49,51,53), were similar between the wild type and the fliL mutant, as confirmed by Western blotting (Fig. 2).…”

Section: Resultsmentioning

confidence: 58%

“…The wavelike morphology of B. burgdorferi, which is dictated at least in part by the presence of flagellar filaments, was not altered in ⌬fliL cells (Fig. 2) (9,12,20,38,43,51,53). Cryo-ET of B. burgdorferi ⌬fliL cells indicated that the periplasmic flagella are connected to the hook-basal body (Fig.…”

Section: Discussionmentioning

confidence: 99%

A Novel Gene Inactivation System Reveals Altered Periplasmic Flagellar Orientation in aBorrelia burgdorferi fliLMutant

et al. 2011

View full text Add to dashboard Cite

Motility and chemotaxis are essential components of pathogenesis for many infectious bacteria, includingBorrelia burgdorferi, the causative agent of Lyme disease. Motility and chemotaxis genes comprise 5 to 6% of the genome of B. burgdorferi, yet the functions of most of those genes remain uncharacterized, mainly due to the paucity of a nonpolar gene inactivation system. In this communication, we describe the development of a novel gene inactivation methodology to target B. burgdorferi fliL, a putative periplasmic flagellar gene located in a large motility operon and transcribed by RNA polymerase containing 70 . Although the morphology of nonpolar fliL mutant cells was indistinguishable from that of wild-type cells, the mutant exhibited a defectivemotility phenotype. Cryo-electron tomography (cryo-ET) of intact organisms revealed that the periplasmic flagella in the fliL mutant were frequently tilted toward the cell pole instead of their normal orientation toward the cell body. These defects were corrected when the mutant was complemented in cis. Moreover, a comparative analysis of flagellar motors from the wild type and the mutant provides the first structural evidence that FliL is localized between the stator and rotor. Our results suggest that FliL is likely involved in coordinating or regulating the orientation of periplasmic flagella in B. burgdorferi.Borrelia burgdorferi is the causative agent of Lyme disease and belongs to a group of bacteria called spirochetes. B. burgdorferi cells have a characteristic flat-wave morphology and unique means of motility (9,10,19,35). As a result of its unique morphology and motility, B. burgdorferi is able to traverse viscous gel-like media in which most other flagellated bacteria slow down or stop (30). Consequently, B. burgdorferi may efficiently bore through host tissues, leading to pathogenesis in the joints, nervous system, and heart (19,30,54). The motility of B. burgdorferi results from the coordinated rotation of the periplasmic flagella residing between the outer membrane and the cell cylinder (9,10,12,20,33,35). The current swimming model suggests that a run occurs when the anterior periplasmic flagella rotate in one direction (e.g., counterclockwise [CCW]) and the posterior flagella rotate in the opposite direction (clockwise [CW]). Reversals occur when periplasmic flagella at both poles of the cell change their direction of rotation. During a nontranslational mode (flex), the periplasmic flagella at both cell poles rotate in the same direction (9,20,35).Bacterial flagella are composed of three major parts: the motor, hook, and filament. The motor can be divided into two functional units, the rotor and the stator. The stator is the torque generator consisting of the MotA-MotB complex. The rotor is composed, minimally, of the MS ring (FliF), the rod, and the switch complex (FliG, FliM, and FliN). In B. burgdorferi, the periplasm-localized flagellar filament consists of the core protein FlaB and the sheath protein FlaA (17, 46). There are 7 to 11 filaments conne...

show abstract

Transformation of Borrelia burgdorferi

2021

Self Cite

View full text Add to dashboard Cite

Transformation techniques used to genetically manipulate Borrelia burgdorferi, the agent of Lyme disease, play a critical role in generating mutants that facilitate analyses of the role of genes in the pathophysiology of this bacterium. A number of borrelial mutants have been successfully isolated and characterized since the first electrotransformation procedure was established 25 years ago (Samuels, 1995). This article is directed at additional considerations for transforming infectious B. burgdorferi to generate strains retaining the plasmid profile of the parental strain, enabling analysis of transformants for in vitro and in vivo phenotypes. These methods are built on previously published protocols and are intended to add steps and tips to enhance transformation efficiency and recovery of strains amenable for studies involving colonization, survival, and transmission of B. burgdorferi during the vector and vertebrate phases of infection.

show abstract

Overexpression of CsrA (BB0184) Alters the Morphology and Antigen Profiles ofBorrelia burgdorferi

Cited by 41 publications

References 89 publications

Oligopeptide Permease A5 Modulates Vertebrate Host-Specific Adaptation of Borrelia burgdorferi

Oligopeptide Permease A5 Modulates Vertebrate Host-Specific Adaptation of Borrelia burgdorferi

A Novel Gene Inactivation System Reveals Altered Periplasmic Flagellar Orientation in aBorrelia burgdorferi fliLMutant

Transformation of Borrelia burgdorferi

Contact Info

Product

Resources

About