Functionality of Sortase A in
            <i>Lactococcus lactis</i>

Dièye, Yakhya; Oxaran, Virginie; Ledue-Clier, Florence; Alkhalaf, W.; Buist, Girbe; Juillard, Vincent; Lee, Chang Won; Piard, Jean-Christophe

doi:10.1128/aem.00928-10

Cited by 39 publications

(50 citation statements)

References 46 publications

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“…Both mutants contain in-frame deletions of approximately 90 % of the sortase gene. Due to the deficiency of SrtA, these strains are also expected to lack the surface display of predicted SDPs (Dieye et al, 2010;Remus et al, 2013). When both DsrtA mutants were examined for their growth characteristics in culture medium (MRS) and survival in SGJ and SIJ, neither L. acidophilus NCK2232 nor L. gasseri NCK2256 showed growth impairment or increased LGAS_1725 1993 2 *The presence of a signal peptide was determined using InterProScan (http://www.ebi.ac.uk/Tools/pfa/iprscan/).…”

Section: Construction Of Sortase-deficient Mutantsmentioning

confidence: 99%

“…Similarly, SDPs play a role in the virulence of several other Gram-positive pathogenic species such as Listeria monocytogenes (Bierne et al, 2002) and Streptococcus pneumoniae (Paterson & Mitchell, 2006). Meanwhile, the role of SrtA has only recently been studied in species of commensal and probiotic microbes, including Lactobacillus casei BL23 (Muñoz-Provencio et al, 2012), Lactococcus lactis IL1403 (Dieye et al, 2010), Lactobacillus plantarum WCFS1 (Remus et al, 2013) and Lactobacillus salivarius UCC118 (O'Callaghan et al, 2012;van Pijkeren et al, 2006). These studies demonstrated that SrtA and SDPs play pivotal roles in the adhesion to intestinal epithelial cells (IECs), immune modulation of DCs, as well as the expression of mucin genes in Caco-2 IECs.…”

Section: Introductionmentioning

confidence: 99%

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Sortase-deficient lactobacilli: effect on immunomodulation and gut retention

et al. 2015

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Surface proteins of probiotic microbes, including Lactobacillus acidophilus and Lactobacillus gasseri, are believed to promote retention in the gut and mediate host-bacterial communications. Sortase, an enzyme that covalently couples a subset of extracellular proteins containing an LPXTG motif to the cell surface, is of particular interest in characterizing bacterial adherence and communication with the mucosal immune system. A sortase gene, srtA, was identified in L. acidophilus NCFM (LBA1244) and L. gasseri ATCC 33323 (LGAS_0825). Additionally, eight and six intact sortase-dependent proteins were predicted in L. acidophilus and L. gasseri, respectively. Due to the role of sortase in coupling these proteins to the cell wall, DsrtA deletion mutants of L. acidophilus and L. gasseri were created using the upp-based counterselective gene replacement system. Inactivation of sortase did not cause significant alteration in growth or survival in simulated gastrointestinal juices. Meanwhile, both DsrtA mutants showed decreased adhesion to porcine mucin in vitro. Murine dendritic cells exposed to the DsrtA mutant of L. acidophilus or L. gasseri induced lower levels of pro-inflammatory cytokines TNF-a and IL-12, respectively, compared with the parent strains. In vivo co-colonization of the L. acidophilus DsrtA mutant and its parent strain in germ-free 129S6/SvEv mice resulted in a significant one-log reduction of the DsrtA mutant population. Additionally, a similar reduction of the DsrtA mutant was observed in the caecum. This study shows for the first time that sortase-dependent proteins contribute to gut retention of probiotic microbes in the gastrointestinal tract.

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Section: Construction Of Sortase-deficient Mutantsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Sortase-deficient lactobacilli: effect on immunomodulation and gut retention

et al. 2015

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“…The transpeptidase sortase A (SrtA) covalently anchors proteins containing an LPXTG motif to the cell wall of a range of Gram-positive bacteria (5)(6)(7)(8)(9). S. epidermidis RP62A expresses 11 proteins containing an LPXTG motif (10).…”

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confidence: 99%

Role for the A Domain of Unprocessed Accumulation-Associated Protein (Aap) in the Attachment Phase of the Staphylococcus epidermidis Biofilm Phenotype

et al. 2014

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e The polysaccharide intercellular adhesin or the cell wall-anchored accumulation-associated protein (Aap) mediates cellular accumulation during Staphylococcus epidermidis biofilm maturation. Mutation of sortase, which anchors up to 11 proteins (including Aap) to the cell wall, blocked biofilm development by the cerebrospinal fluid isolate CSF41498. Aap was implicated in this phenotype when Western blots and two-dimensional (2D) electrophoresis revealed increased levels of the protein in culture supernatants. Unexpectedly, reduced levels of primary attachment were associated with impaired biofilm formation by CSF41498 srtA and aap mutants. In contrast to previous studies, which implicated Aap proteolytic cleavage and, specifically, the Aap B domains in biofilm accumulation, the CSF41498 Aap protein was unprocessed. Furthermore, aap appeared to play a less important role in the biofilm phenotype of S. epidermidis 1457, in which the Aap protein is processed. Anti-Aap A-domain IgG inhibited primary attachment and biofilm formation in strain CSF41498 but not in strain 1457. The nucleotide sequences of the aap gene A-domain region and cleavage site in strains CSF41498 and 1457 were identical, implicating altered protease activity in the differential Aap processing results in the two strains. These data reveal a new role for the A domain of unprocessed Aap in the attachment phase of biofilm formation and suggest that extracellular protease activity can influence whether Aap contributes to the attachment or accumulation phases of the S. epidermidis biofilm phenotype.

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“…This has been reported for sortase mutants of different classes in many Gram-positive microorganisms including LAB. That is the case for the different mutants of L. casei BL23 obtained in this study and also for the srtA deletion mutant in the model LAB L. lactis (Dieye, et al, 2010). Only in the case of Bacillus anthracis class B sortase and under iron-limiting conditions significant growth defects have been detected (Maresso, et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

“…This is in contrast to recent results obtained in L. lactis, where a srtA mutation completely abolished the anchoring to the cell wall of all the sortase substrates tested. Therefore, as reported for many other Grampositive bacteria, srtA is the single gene responsible for the general sortase activity in L. lactis (Dieye, et al, 2010) Also, Dieye and coworkers used two-dimensional gel electrophoresis of surface proteins to search for proteins absent in the L. lactis srtA mutant, and they identified several sortase substrates (Dieye, et al, 2010). However, and in agreement with the results of surface anchoring, our efforts to detect the absence/reduction of surface proteins in the BL23 srtA1/srtA2 mutant by this technique were not successful (data not shown).…”

Section: Discussionmentioning

confidence: 99%

Caracterización de factores de adhesión a proteínas de la matriz extracelular en Lactobacillus casei

Provencio¹

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CERTIFICA:Que la memoria titulada "Caracterización de factores de adhesión a proteínas de la matriz extracelular en Lactobacillus casei" presentada por el licenciado Diego Muñoz Provencio para optar al grado de doctor por la Universidad Politécnica de Valencia, ha sido realizada en el Instituto de Agroquímica y Tecnología de Alimentos (IATA-CSIC) bajo mi dirección y que reúne las condiciones necesarias para ser defendida por su autor ante el tribunal correspondiente. Valencia, Septiembre 2011Fdo.: Dr. Vicente Monedero García I AGRADECIMIENTOS Lo que estáis leyendo no son los agradecimientos originales. Acabaron siendo demasiado buenos, notablemente más largos de lo normal y de una escandalosa incorrección política. Por el momento duermen el sueño de los justos en un cajón y me los guardo como apuntes para una novela.Agradezco a la Generalitat Valenciana que tuviera a bien otorgarme una beca FPI y al Instituto Danone la concesión de una beca de investigación sin las cuales hubiera debido hacer la tesis simplemente por amor al arte y viviendo debajo de un puente. Mi familia tiene mis agradecimientos por descontado, son mi grupo de apoyo, me impulsan a seguir.Sin Vicente, mi director de tesis, nada de esto hubiera sido posible (GRACIAS con mayúsculas). Gracias también a mis tutores, Javier Buesa y Manuel Hernández, pese a que solo les vi fugazmente sé que me hubieran ayudado de haberlo requerido. Gracias a Gaspar que fue el primero con el que me puse en contacto y que inició la cadena de acontecimientos que me llevaron a este laboratorio.Del periodo neurocientífico y cinematográfico debo dar las gracias a Víctor Meseguer, un amigo de los que siempre están cuando los necesitas, a Rodolfo Madrid que siempre se portó muy bien conmigo y a Neus Ayuso que me acogió en su casa como a alguien de su familia.Agradezco a todo el personal del IATA el buen trato recibido. En especial aprecio mucho el momento "¡Hola, hola!" diario de Emilia y las conversaciones con Fran, Miguel y Alí.De los mandamases no citados ya en el primer párrafo, debo dar las gracias a Mª Jesús por darme esos buenos kilos de limones cada año y a Manolo por ayudarme a poner en hora el reloj con su puntualidad britanica.Le doy las gracias a Juan, Luisa, Sneyder, Adriana, Christine y Carmen que me permitieron adaptarme fácilmente al lugar al empezar la tesis. IIChristine se merece una mención especial; es con la que más tiempo he coincidido y me ha ayudado siempre que me ha hecho falta.A Lourdes y Maite les agradezco su facilidad para transmitir alegría, a Pepe lo fácil que es llevarse bien con él y a Ahmed que siempre se acuerde de mí cuando vuelve en su peregrinaje intermitente. A Inma, Amparo, María y Pascual les agradezco el que siempre me recibieran con una sonrisa.A Cristina, Chema, Ainhoa y Amalia les doy las gracias por constituir el ecosistema único del laboratorio 007. A los dos primeros por sus consejos y buen humor, a la tercera por sus ratos buenos y a la cuarta por ser como mi mami en el laboratorio.Gracias a Jesús por intentar lo del Biacore c...

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Functionality of Sortase A in Lactococcus lactis

Cited by 39 publications

References 46 publications

Sortase-deficient lactobacilli: effect on immunomodulation and gut retention

Sortase-deficient lactobacilli: effect on immunomodulation and gut retention

Role for the A Domain of Unprocessed Accumulation-Associated Protein (Aap) in the Attachment Phase of the Staphylococcus epidermidis Biofilm Phenotype

Caracterización de factores de adhesión a proteínas de la matriz extracelular en Lactobacillus casei

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