scholarly journals 1P183 Structural analysis of the periplasmic flagella in Borrelia burgdorferi

2005 ◽  
Vol 45 (supplement) ◽  
pp. S77
Author(s):  
S. Shibata ◽  
S. Aizawa ◽  
Nyles W. Charon
Keyword(s):  
Structural Analysis ◽  
Borrelia Burgdorferi ◽  
Periplasmic Flagella

scholarly journals The Borrelia burgdorferi 37-Kilodalton Immunoblot Band (P37) Used in Serodiagnosis of Early Lyme Disease Is the flaA Gene Product

1999 ◽  
Vol 37 (3) ◽  
pp. 548-552 ◽  
Author(s):  
Robert D. Gilmore ◽  
Rendi L. Murphree ◽  
Angela M. James ◽  
Sarah A. Sullivan ◽  
Barbara J. B. Johnson
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Genomic Library ◽  
Disease Patient ◽  
Humoral Response ◽  
Immunoglobulin M ◽  
Leader Peptide ◽  
Serum Samples ◽  
Present Generation ◽  
Periplasmic Flagella

The 37-kDa protein (P37) of Borrelia burgdorferi is an antigen that elicits an early immunoglobulin M (IgM) antibody response in Lyme disease patients. The P37 gene was cloned from aB. burgdorferi genomic library by screening with antibody from a Lyme disease patient who had developed a prominent humoral response to the P37 antigen. DNA sequence analysis of this clone revealed the identity of P37 to be FlaA, an outer sheath protein of the periplasmic flagella. Recombinant P37 expression was accomplished inEscherichia coli by using a gene construct with the leader peptide deleted and fused to a 38-kDa E. coli protein. The recombinant antigen was reactive in IgM immunoblots using serum samples from patients clinically diagnosed with early Lyme disease that had been scored positive for B. burgdorferi anti-P37 reactivity. Lyme disease patient samples serologically negative for theB. burgdorferi P37 protein did not react with the recombinant. Recombinant P37 may be a useful component of a set of defined antigens for the serodiagnosis of early Lyme disease. This protein can be utilized as a marker in diagnostic immunoblots, aiding in the standardization of the present generation of IgM serologic tests.

scholarly journals BB0259 Encompasses a Peptidoglycan Lytic Enzyme Function for Proper Assembly of Periplasmic Flagella in Borrelia burgdorferi

2021 ◽  
Vol 12 ◽  
Author(s):  
Hui Xu ◽  
Bo Hu ◽  
David A. Flesher ◽  
Jun Liu ◽  
Md A. Motaleb
Keyword(s):  
Borrelia Burgdorferi ◽  
Electron Tomography ◽  
Lytic Enzyme ◽  
Enzyme Protein ◽  
Lytic Transglycosylase ◽  
Filament Assembly ◽  
Cryo Electron Tomography ◽  
Mutant Strains ◽  
Periplasmic Flagella ◽  
Dual Domain

Assembly of the bacterial flagellar rod, hook, and filament requires penetration through the peptidoglycan (PG) sacculus and outer membrane. In most β- and γ-proteobacteria, the protein FlgJ has two functional domains that enable PG hydrolyzing activity to create pores, facilitating proper assembly of the flagellar rod. However, two distinct proteins performing the same functions as the dual-domain FlgJ are proposed in δ- and ε-proteobacteria as well as spirochetes. The Lyme disease spirochete Borrelia burgdorferi genome possesses a FlgJ and a PG lytic SLT enzyme protein homolog (BB0259). FlgJ in B. burgdorferi is crucial for flagellar hook and filament assembly but not for the proper rod assembly reported in other bacteria. However, BB0259 has never been characterized. Here, we use cryo-electron tomography to visualize periplasmic flagella in different bb0259 mutant strains and provide evidence that the E580 residue of BB0259 is essential for PG-hydrolyzing activity. Without the enzyme activity, the flagellar hook fails to penetrate through the pores in the cell wall to complete assembly of an intact periplasmic flagellum. Given that FlgJ and BB0259 interact with each other, they likely coordinate the penetration through the PG sacculus and assembly of a functional flagellum in B. burgdorferi and other spirochetes. Because of its role, we renamed BB0259 as flagellar-specific lytic transglycosylase or LTaseBb.

scholarly journals FlhF regulates the number and configuration of periplasmic flagella in Borrelia burgdorferi

2020 ◽  
Vol 113 (6) ◽  
pp. 1122-1139 ◽  
Author(s):  
Kai Zhang ◽  
Jun He ◽  
Claudio Catalano ◽  
Youzhong Guo ◽  
Jun Liu ◽  
...  
Keyword(s):  
Borrelia Burgdorferi ◽  
Periplasmic Flagella

scholarly journals Borrelia burgdorferi Uniquely Regulates Its Motility Genes and Has an Intricate Flagellar Hook-Basal Body Structure

2008 ◽  
Vol 190 (6) ◽  
pp. 1912-1921 ◽  
Author(s):  
Melanie S. Sal ◽  
Chunhao Li ◽  
M. A. Motalab ◽  
Satoshi Shibata ◽  
Shin-Ichi Aizawa ◽  
...  
Keyword(s):  
Borrelia Burgdorferi ◽  
Gene Transcription ◽  
Basal Body ◽  
Transcriptional Control ◽  
Targeted Mutagenesis ◽  
Gene Encoding ◽  
High Molecular Weight Complex ◽  
Promoter Recognition ◽  
Flagellar Filament ◽  
Periplasmic Flagella

ABSTRACT Borrelia burgdorferi is a flat-wave, motile spirochete that causes Lyme disease. Motility is provided by periplasmic flagella (PFs) located between the cell cylinder and an outer membrane sheath. The structure of these PFs, which are composed of a basal body, a hook, and a filament, is similar to the structure of flagella of other bacteria. To determine if hook formation influences flagellin gene transcription in B. burgdorferi, we inactivated the hook structural gene flgE by targeted mutagenesis. In many bacteria, completion of the hook structure serves as a checkpoint for transcriptional control of flagellum synthesis and other chemotaxis and motility genes. Specifically, the hook allows secretion of the anti-sigma factor FlgM and concomitant late gene transcription promoted by σ28. However, the control of B. burgdorferi PF synthesis differs from the control of flagellum synthesis in other bacteria; the gene encoding σ28 is not present in the genome of B. burgdorferi, nor are any σ28 promoter recognition sequences associated with the motility genes. We found that B. burgdorferi flgE mutants lacked PFs, were rod shaped, and were nonmotile, which substantiates previous evidence that PFs are involved in both cell morphology and motility. Although most motility and chemotaxis gene products accumulated at wild-type levels in the absence of FlgE, mutant cells had markedly decreased levels of the flagellar filament proteins FlaA and FlaB. Further analyses showed that the reduction in the levels of flagellin proteins in the spirochetes lacking FlgE was mediated at the posttranscriptional level. Taken together, our results indicate that in B. burgdorferi, the completion of the hook does not serve as a checkpoint for transcriptional regulation of flagellum synthesis. In addition, we also present evidence that the hook protein in B. burgdorferi forms a high-molecular-weight complex and that formation of this complex occurs in the periplasmic space.

scholarly journals Structure and Expression of the FlaA Periplasmic Flagellar Protein of Borrelia burgdorferi

1998 ◽  
Vol 180 (9) ◽  
pp. 2418-2425 ◽  
Author(s):  
Yigong Ge ◽  
Chunhao Li ◽  
Linda Corum ◽  
Clive A. Slaughter ◽  
Nyles W. Charon
Keyword(s):  
Borrelia Burgdorferi ◽  
Treponema Pallidum ◽  
The Other ◽  
Wild Type ◽  
Serpulina Hyodysenteriae ◽  
Flagellar Proteins ◽  
Flagellar Protein ◽  
Periplasmic Flagella ◽  
Triton X ◽  
Translational Level

ABSTRACT The spirochete which causes Lyme disease, Borrelia burgdorferi, has many features common to other spirochete species. Outermost is a membrane sheath, and within this sheath are the cell cylinder and periplasmic flagella (PFs). The PFs are subterminally attached to the cell cylinder and overlap in the center of the cell. Most descriptions of the B. burgdorferi flagellar filaments indicate that these organelles consist of only one flagellin protein (FlaB). In contrast, the PFs from other spirochete species are comprised of an outer layer of FlaA and a core of FlaB. We recently found that a flaA homolog was expressed in B. burgdorferi and that it mapped in a fla/che operon. These results led us to analyze the PFs and FlaA of B. burgdorferi in detail. Using Triton X-100 to remove the outer membrane and isolate the PFs, we found that the 38.0-kDa FlaA protein purified with the PFs in association with the 41.0-kDa FlaB protein. On the other hand, purifying the PFs by using Sarkosyl resulted in no FlaA in the isolated PFs. Sarkosyl has been used by others to purifyB. burgdorferi PFs, and our results explain in part their failure to find FlaA. Unlike other spirochetes, B. burgdorferi FlaA was expressed at a lower level than FlaB. In characterizing FlaA, we found that it was posttranslationally modified by glycosylation, and thus it resembles its counterpart fromSerpulina hyodysenteriae. We also tested if FlaA was synthesized in a spontaneously occurring PF mutant of B. burgdorferi (HB19Fla−). Although this mutant still synthesizedflaA message in amounts similar to the wild-type amounts, it failed to synthesize FlaA protein. These results suggest that, in agreement with data found for FlaB and other spirochete flagellar proteins, FlaA is likely to be regulated on the translational level. Western blot analysis using Treponema pallidum anti-FlaA serum indicated that FlaA was antigenically well conserved in several spirochete species. Taken together, the results indicate that both FlaA and FlaB comprise the PFs of B. burgdorferi and that they are regulated differently from flagellin proteins of other bacteria.

scholarly journals Motility Is Crucial for the Infectious Life Cycle of Borrelia burgdorferi

2013 ◽  
Vol 81 (6) ◽  
pp. 2012-2021 ◽  
Author(s):  
Syed Z. Sultan ◽  
Akarsh Manne ◽  
Philip E. Stewart ◽  
Aaron Bestor ◽  
Patricia A. Rosa ◽  
...  
Keyword(s):  
Life Cycle ◽  
Borrelia Burgdorferi ◽  
Mammalian Host ◽  
Major Protein ◽  
Content Type ◽  
Absolute Requirement ◽  
Wave Morphology ◽  
Minor Protein ◽  
Periplasmic Flagella ◽  
First Time

ABSTRACTThe Lyme disease spirochete,Borrelia burgdorferi, exists in a zoonotic cycle involving an arthropod tick and mammalian host. Dissemination of the organism within and between these hosts depends upon the spirochete's ability to traverse through complex tissues. Additionally, the spirochete outruns the host immune cells while migrating through the dermis, suggesting the importance ofB. burgdorferimotility in evading host clearance.B. burgdorferi's periplasmic flagellar filaments are composed primarily of a major protein, FlaB, and minor protein, FlaA. By constructing aflaBmutant that is nonmotile, we investigated for the first time the absolute requirement for motility in the mouse-tick life cycle ofB. burgdorferi. We found that whereas wild-type cells are motile and have a flat-wave morphology, mutant cells were nonmotile and rod shaped. These mutants were unable to establish infection in C3H/HeN mice via either needle injection or tick bite. In addition, these mutants had decreased viability in fed ticks. Our studies provide substantial evidence that the periplasmic flagella, and consequently motility, are critical not only for optimal survival in ticks but also for infection of the mammalian host by the arthropod tick vector.

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