Toolbox of Molecular Techniques for Studying Leptospira Spp.

Picardeau, Mathieu

doi:10.1007/82_2017_45

Cited by 18 publications

(16 citation statements)

References 114 publications

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“…Taken together, these data are consistent with FlaB1-4 forming the filament core, while FcpA, possibly in tandem with FlaA1 and FlaA2, is a major component of the sheath. Conservation of the flagellar components in saprophytic and pathogenic Leptospira strains (Fouts et al, 2016 ) makes of the saprophyte L. biflexa , an ideal model to understand spirochete motility, highlighting that this species is relatively easy for in vitro culturing and genetic manipulations compared to pathogenic species (Picardeau, 2017a ). In this study, following the screening of a library of random mutants in L. biflexa , we identified and characterized a novel component of the flagellar filament of Leptospira .…”

Section: Discussionmentioning

confidence: 99%

“…In Leptospira one PF is attached at each pole of the cell, extending axially toward the center without overlapping (Picardeau, 2017b ). The Leptospira genus comprises pathogenic and non-pathogenic species, which are poorly transformable bacteria (Picardeau, 2017a ). Nevertheless, several non-motile mutants were described in the last few years by screening a library of random transposon mutants (Lambert et al, 2012 ), isolation of spontaneous mutants (Fontana et al, 2016 ; Wunder et al, 2016b ), and targeted mutagenesis (Picardeau et al, 2001 ; Liao et al, 2009 ).…”

Section: Introductionmentioning

confidence: 99%

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FcpB Is a Surface Filament Protein of the Endoflagellum Required for the Motility of the Spirochete Leptospira

Wunder

Slamti

Suwondo

et al. 2018

Front. Cell. Infect. Microbiol.

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The spirochete endoflagellum is a unique motility apparatus among bacteria. Despite its critical importance for pathogenesis, the full composition of the flagellum remains to be determined. We have recently reported that FcpA is a novel flagellar protein and a major component of the sheath of the filament of the spirochete Leptospira. By screening a library of random transposon mutants in the spirochete Leptospira biflexa, we found a motility-deficient mutant harboring a disruption in a hypothetical gene of unknown function. Here, we show that this gene encodes a surface component of the endoflagellar filament and is required for typical hook- and spiral-shaped ends of the cell body, coiled structure of the endoflagella, and high velocity phenotype. We therefore named the gene fcpB for flagellar-coiling protein B. fcpB is conserved in all members of the Leptospira genus, but not present in other organisms including other spirochetes. Complementation of the fcpB− mutant restored the wild-type morphology and motility phenotypes. Immunoblotting with anti-FcpA and anti-FcpB antisera and cryo-electron microscopy of the filament indicated that FcpB assembled onto the surface of the sheath of the filament and mostly located on the outer (convex) side of the coiled filament. We provide evidence that FcpB, together with FcpA, are Leptospira-specific novel components of the sheath of the filament, key determinants of the coiled and asymmetric structure of the endoflagella and are essential for high velocity. Defining the components of the endoflagella and their functions in these atypical bacteria should greatly enhance our understanding of the mechanisms by which these bacteria produce motility.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

FcpB Is a Surface Filament Protein of the Endoflagellum Required for the Motility of the Spirochete Leptospira

Wunder

Slamti

Suwondo

et al. 2018

Front. Cell. Infect. Microbiol.

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“…Genetic manipulation is a strategy used to study the biology of pathogenic microorganisms and their virulence factors, which are important in host-pathogen interactions. The number of genetic tool available for Leptospira has increased over the years but is still limited when compared to other pathogenic bacteria [ 29 ].…”

Section: Discussionmentioning

confidence: 99%

Heterologous Expression of the Pathogen-Specific LIC11711 Gene in the Saprophyte L. biflexa Increases Bacterial Binding to Laminin and Plasminogen

2020

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Leptospirosis is a febrile disease and the etiological agents are pathogenic bacteria of the genus Leptospira. The leptospiral virulence mechanisms are not fully understood and the application of genetic tools is still limited, despite advances in molecular biology techniques. The leptospiral recombinant protein LIC11711 has shown interaction with several host components, indicating a potential function in virulence. This study describes a system for heterologous expression of the L. interrogans gene lic11711 using the saprophyte L. biflexa serovar Patoc as a surrogate, aiming to investigate its possible activity in bacterial virulence. Heterologous expression of LIC11711 was performed using the pMaOri vector under regulation of the lipL32 promoter. The protein was found mainly on the leptospiral outer surface, confirming its location. The lipL32 promoter enhanced the expression of LIC11711 in L. biflexa compared to the pathogenic strain, indicating that this strategy may be used to overexpress low-copy proteins. The presence of LIC11711 enhanced the capacity of L. biflexa to adhere to laminin (Lam) and plasminogen (Plg)/plasmin (Pla) in vitro, suggesting the involvement of this protein in bacterial pathogenesis. We show for the first time that the expression of LIC11711 protein of L. interrogans confers a virulence-associated phenotype on L. biflexa, pointing out possible mechanisms used by pathogenic leptospires.

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“…Molecular techniques for the genetic analysis of Leptospira remain very limited [12]. Mutants, a few plasmids, and only three phages are available for a few model strains.…”

Section: Introductionmentioning

confidence: 99%

Transposable Prophages in Leptospira: An Ancient, Now Diverse, Group Predominant in Causative Agents of Weil’s Disease

Ndela

Enault

Toussaint

2021

IJMS

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The virome associated with the corkscrew shaped bacterium Leptospira, responsible for Weil’s disease, is scarcely known, and genetic tools available for these bacteria remain limited. To reduce these two issues, potential transposable prophages were searched in Leptospiraceae genomes. The 236 predicted transposable prophages were particularly abundant in the most pathogenic leptospiral clade, being potentially involved in the acquisition of virulent traits. According to genomic similarities and phylogenies, these prophages are distantly related to known transposable phages and are organized into six groups, one of them encompassing prophages with unusual TA-TA ends. Interestingly, structural and transposition proteins reconstruct different relationships between groups, suggesting ancestral recombinations. Based on the baseplate phylogeny, two large clades emerge, with specific gene-contents and high sequence divergence reflecting their ancient origin. Despite their high divergence, the size and overall genomic organization of all prophages are very conserved, a testimony to the highly constrained nature of their genomes. Finally, similarities between these prophages and the three known non-transposable phages infecting L. biflexa, suggest gene transfer between different Caudovirales inside their leptospiral host, and the possibility to use some of the transposable prophages in that model strain.

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Toolbox of Molecular Techniques for Studying Leptospira Spp.

Cited by 18 publications

References 114 publications

FcpB Is a Surface Filament Protein of the Endoflagellum Required for the Motility of the Spirochete Leptospira

FcpB Is a Surface Filament Protein of the Endoflagellum Required for the Motility of the Spirochete Leptospira

Heterologous Expression of the Pathogen-Specific LIC11711 Gene in the Saprophyte L. biflexa Increases Bacterial Binding to Laminin and Plasminogen

Transposable Prophages in Leptospira: An Ancient, Now Diverse, Group Predominant in Causative Agents of Weil’s Disease

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