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1.
Mol Microbiol ; 118(5): 503-509, 2022 11.
Artículo en Inglés | MEDLINE | ID: mdl-36203248

RESUMEN

LeuO is a LysR-type transcriptional regulator in bacteria. It determines the regulation of numerous genes related to stress response and virulence. Thus, four exciting areas of research are discussed herein. One pertains the leuO gene, which in S. Typhi and in E. coli contains multiple forward promoters as well as reverse promoters, even though it is expressed at very low levels, that is, it is quiescent. Such multiplicity might allow for a greater plasticity in regulation, or even aid in maintaining the quiescence, in processes that appear to involve many nucleoid-associated proteins in a second area of opportunity. A third one relates to the effector-binding domain of the LeuO regulator, which is highly conserved in S. enterica and in E. coli and determines its activity as a regulator of transcription. A fourth area regards the role of the CRISPR-Cas system in gene regulation in S. Typhi; a system that is regulated by LeuO.


Asunto(s)
Proteínas de Escherichia coli , Regulación Bacteriana de la Expresión Génica , Regulación Bacteriana de la Expresión Génica/genética , Factores de Transcripción/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/metabolismo , Sistemas CRISPR-Cas/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo
2.
Microbiology (Reading) ; 163(2): 253-265, 2017 02.
Artículo en Inglés | MEDLINE | ID: mdl-28270274

RESUMEN

The CRISPR-Cas system is involved in bacterial immunity, virulence, gene regulation, biofilm formation and sporulation. In Salmonella enterica serovar Typhi, this system consists of five transcriptional units including antisense RNAs. It was determined that these genetic elements are expressed in minimal medium and are up-regulated by pH. In addition, a transcriptional characterization of cas3 and ascse2-1 is included herein.


Asunto(s)
Proteínas Asociadas a CRISPR/genética , Sistemas CRISPR-Cas/genética , ADN Helicasas/genética , Regulación Bacteriana de la Expresión Génica/genética , ARN sin Sentido/genética , Salmonella typhi/genética , Cloranfenicol O-Acetiltransferasa/metabolismo , Ensayo de Cambio de Movilidad Electroforética , Transcripción Genética/genética , Activación Transcripcional/genética , Regulación hacia Arriba/genética
3.
BMC Microbiol ; 16: 18, 2016 Feb 09.
Artículo en Inglés | MEDLINE | ID: mdl-26862033

RESUMEN

BACKGROUND: Classical strains of Salmonella enterica serovar Typhimurium (Typhimurium) predominantly cause a self-limiting diarrheal illness in humans and a systemic disease in mice. In this study, we report the characterization of a strain isolated from a blood-culture taken from a 15-year old woman suffering from invasive severe salmonellosis, refractory to conventional therapy with extended-spectrum cephalosporin (ESC). RESULTS: The strain, named 33676, was characterized as multidrug-resistant Salmonella serogroup A by biochemical, antimicrobial and serological tests. Multilocus sequence typing (MLST) and XbaI macrorestrictions (PFGE) showed that strain 33676 belonged to the Typhimurium ST213 genotype, previously described for other Mexican Typhimurium strains. PCR analyses revealed the presence of IncA/C, IncFIIA and ColE1-like plasmids and the absence of the Salmonella virulence plasmid (pSTV). Conjugation assays showed that the ESC-resistance gene bla CMY-2 was carried on the conjugative IncF plasmid, instead of the IncA/C plasmid, as found in previously studied ST213 strains. Although the IncA/C plasmid conferred most of the observed antimicrobial resistances it was not self-conjugative; it was rather able to conjugate by co-integrating with the IncF plasmid. Strain 33676 was fully attenuated for virulence in BALB/c mice infections. Both type-three secretion system (T3SS), encoded in Salmonella pathogenicity islands 1 and 2 (SPI-1 and SPI-2), were functional in the 33676 strain and, interestingly, this strain produced the H2 FljB flagellin instead of the H1 FliC flagellin commonly expressed by S. enterica strains. CONCLUSIONS: Strain 33676 showed two main features that differentiate it from the originally described ST213 strains: 1) the bla CMY-2 gene was not carried on the IncA/C plasmid, but on a conjugative IncF plasmid, which may open a new route of dissemination for this ESC-resistance gene, and 2) it expresses the H2 FljB flagella, in contrast with the other ST213 and most Typhimurium reference strains. To our knowledge this is the first report of an IncF bla CMY-2-carrying plasmid in Salmonella.


Asunto(s)
Farmacorresistencia Bacteriana Múltiple , Plásmidos/genética , Infecciones por Salmonella/microbiología , Salmonella typhimurium/enzimología , Salmonella typhimurium/patogenicidad , beta-Lactamasas/metabolismo , Animales , Antibacterianos/farmacología , Femenino , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Plásmidos/metabolismo , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genética , Virulencia , beta-Lactamasas/genética
4.
Mol Microbiol ; 91(6): 1054-6, 2014 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-24405400

RESUMEN

A new mechanism for the turning-off of gene expression in Salmonella Pathogenicity Island 1 (SPI-1) is proposed by Espinosa and Casadesús, which involves the action of the LeuO quiescent regulator, by two different pathways. A major one through the activation of the hilE gene, where the HilE protein would in turn inactivate HilD, a major positive transcriptional regulator of SPI-1; and a minor HilE-HilD-independent pathway. This could constitute a back-up or an aid for the turn-off of SPI-1 genes mediated by the nucleoid protein H-NS.


Asunto(s)
Proteínas Bacterianas/biosíntesis , Regulación Bacteriana de la Expresión Génica , Islas Genómicas , Proteínas Represoras/biosíntesis , Salmonella typhimurium/genética , Factores de Transcripción/metabolismo , Factores de Virulencia/genética
5.
Microbiology (Reading) ; 161(Pt 4): 903-13, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25667010

RESUMEN

RcsC is a hybrid histidine kinase that forms part of a phospho-relay signal transduction pathway with RcsD and RcsB. Besides the typical domains of a sensor kinase, i.e. the periplasmic (P), linker (L), dimerization and H-containing (A), and ATP-binding (B) domains, RcsC possesses a receiver domain (D) at the carboxy-terminal domain. To study the role played by each of the RcsC domains, four plasmids containing several of these domains were constructed (PLAB, LAB, AB and ABD) and transformed into Escherichia coli K-12 strain BW25113. Different amounts of biofilm were produced, depending on the RcsC domains expressed: the plasmid expressing the ABD subdomains produced the highest amount of biofilm. This phenotype was also observed when the plasmids were transformed in a ΔrcsCDB strain. Biofilm formation was abolished in the pgaABCD and nhaR backgrounds. The results indicate the existence of a novel signalling pathway that depends on RcsC, yet independent of RcsD and RcsB, that activates the pgaABCD operon and, as a consequence, biofilm formation. This signalling pathway involves the secondary metabolite acetyl phosphate and the response regulator OmpR.


Asunto(s)
Acetilglucosamina/biosíntesis , Biopelículas , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Escherichia coli/fisiología , Eliminación de Gen , Complejos Multienzimáticos/genética , Complejos Multienzimáticos/metabolismo , Fosfoproteínas Fosfatasas/genética , Fosfoproteínas Fosfatasas/metabolismo , Proteínas Quinasas/genética , Proteínas Quinasas/metabolismo , Transducción de Señal , Activación Enzimática , Proteínas de Escherichia coli/química , Complejos Multienzimáticos/química , Operón , Fosfoproteínas Fosfatasas/química , Monoéster Fosfórico Hidrolasas/metabolismo , Regiones Promotoras Genéticas , Unión Proteica , Dominios y Motivos de Interacción de Proteínas , Proteínas Quinasas/química , Proteínas Recombinantes de Fusión
6.
J Bacteriol ; 196(12): 2143-54, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24659766

RESUMEN

LeuO is a LysR-type transcriptional regulator (LTTR) that has been described to be a global regulator in Escherichia coli and Salmonella enterica, since it positively and negatively regulates the expression of genes involved in multiple biological processes. LeuO is comprised of an N-terminal DNA-binding domain (DBD) with a winged helix-turn-helix (wHTH) motif and of a long linker helix (LH) involved in dimerization that connects the DBD with the C-terminal effector-binding domain (EBD) or regulatory domain (RD; which comprises subdomains RD-I and RD-II). Here we show that the oligomeric structure of LeuO is a tetramer that binds with high affinity to DNA. A collection of single amino acid substitutions in the LeuO DBD indicated that this region is involved in oligomerization, in positive and negative regulation, as well as in DNA binding. Mutants with point mutations in the central and C-terminal regions of RD-I were affected in transcriptional activation. Deletion of the RD-II and RD-I C-terminal subdomains affected not only oligomerization but also DNA interaction, showing that they are involved in positive and negative regulation. Together, these data demonstrate that not only the C terminus but also the DBD of LeuO is involved in oligomer formation; therefore, each LeuO domain appears to act synergistically to maintain its regulatory functions in Salmonella enterica serovar Typhi.


Asunto(s)
Proteínas Bacterianas/metabolismo , ADN Bacteriano/metabolismo , Regulación Bacteriana de la Expresión Génica/fisiología , Salmonella typhi/metabolismo , Transcripción Genética , Secuencia de Aminoácidos , Proteínas Bacterianas/genética , Eliminación de Gen , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Unión Proteica , Conformación Proteica , Salmonella typhi/genética
7.
Microbiology (Reading) ; 160(Pt 4): 733-741, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24440835

RESUMEN

The Salmonella enterica ompS1 gene encodes a quiescent porin that belongs to the OmpC/OmpF family. In the present work we analysed the regulatory effects of OmpR phosphorylation on ompS1 expression. We found that in vivo, OmpR in its phosphorylated form (OmpR-P) was important in the regulation of the two ompS1 promoters: OmpR-P activated the P1 promoter and repressed the P2 promoter in an EnvZ-dependent manner; expression occurs from the P2 promoter in an ompR mutant. In vitro, OmpR-P had a higher DNA-binding-affinity to the ompS1 promoter region than OmpR and OmpRD55A, showing an affinity even higher than that of equivalent DNA regions in the 5'-upstream regulatory sequence of the major porin-encoding genes ompC and ompF. By analysing different environmental conditions, we found that glucose and glycerol enhanced ompS1 expression in the wild-type strain. Interestingly the stimulation by glycerol was OmpR-dependent while the effect of glucose was still observed in the absence of OmpR. Acetyl phosphate produced by the AckA-Pta pathway did not influence ompS1 regulation. These data indicate the important role of the phosphorylation in the activity of OmpR on the differential regulation of both ompS1 promoters and its impact on the pathogenesis.


Asunto(s)
Proteínas de la Membrana Bacteriana Externa/biosíntesis , Regulación Bacteriana de la Expresión Génica , Porinas/biosíntesis , Regiones Promotoras Genéticas , Procesamiento Proteico-Postraduccional , Salmonella typhi/genética , Salmonella typhi/metabolismo , Factores de Transcripción/metabolismo , Expresión Génica/efectos de los fármacos , Glucosa/metabolismo , Glicerol/metabolismo , Fosforilación , Unión Proteica
8.
Immunology ; 139(4): 459-71, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23432484

RESUMEN

Salmonella enterica serovar Typhi (S. Typhi) is the causal agent of typhoid fever, a disease that primarily affects developing countries. Various antigens from this bacterium have been reported to be targets of the immune response. Recently, the S. Typhi genome has been shown to encode two porins--OmpS1 and OmpS2--which are expressed at low levels under in vitro culture conditions. In this study, we demonstrate that immunizing mice with either OmpS1 or OmpS2 induced production of specific, long-term antibody titres and conferred protection against S. Typhi challenge; in particular, OmpS1 was more immunogenic and conferred greater protective effects than OmpS2. We also found that OmpS1 is a Toll-like receptor 4 (TLR4) agonist, whereas OmpS2 is a TLR2 and TLR4 agonist. Both porins induced the production of tumour necrosis factor and interleukin-6, and OmpS2 was also able to induce interleukin-10 production. Furthermore, OmpS1 induced the over-expression of MHC II molecules in dendritic cells and OmpS2 induced the over-expression of CD40 molecules in macrophages and dendritic cells. Co-immunization of OmpS1 or OmpS2 with ovalbumin (OVA) increased anti-OVA antibody titres, the duration and isotype diversity of the OVA-specific antibody response, and the proliferation of T lymphocytes. These porins also had adjuvant effects on the antibody response when co-immunized with either the Vi capsular antigen from S. Typhi or inactivated 2009 pandemic influenza A(H1N1) virus [A(H1N1)pdm09]. Taken together, the data indicate that OmpS1 and OmpS2, despite being expressed at low levels under in vitro culture conditions, are potent protective immunogens with intrinsic adjuvant properties.


Asunto(s)
Adyuvantes Inmunológicos , Anticuerpos Antibacterianos/sangre , Proteínas de la Membrana Bacteriana Externa/inmunología , Porinas/inmunología , Vacunas contra la Salmonella/inmunología , Salmonella typhi/inmunología , Fiebre Tifoidea/prevención & control , Adyuvantes Inmunológicos/administración & dosificación , Adyuvantes Inmunológicos/genética , Animales , Proteínas de la Membrana Bacteriana Externa/administración & dosificación , Proteínas de la Membrana Bacteriana Externa/genética , Células Dendríticas/inmunología , Relación Dosis-Respuesta a Droga , Femenino , Células HEK293 , Antígenos de Histocompatibilidad Clase II/metabolismo , Humanos , Inmunización , Subtipo H1N1 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/inmunología , Interleucina-10/metabolismo , Interleucina-6/metabolismo , Activación de Linfocitos , Macrófagos/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Transgénicos , Ovalbúmina/inmunología , Polisacáridos Bacterianos/inmunología , Porinas/administración & dosificación , Porinas/genética , Vacunas contra la Salmonella/administración & dosificación , Vacunas contra la Salmonella/genética , Salmonella typhi/genética , Linfocitos T/inmunología , Factores de Tiempo , Receptor Toll-Like 2/agonistas , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 4/agonistas , Receptor Toll-Like 4/metabolismo , Transfección , Factor de Necrosis Tumoral alfa/metabolismo , Fiebre Tifoidea/sangre , Fiebre Tifoidea/inmunología , Fiebre Tifoidea/microbiología
9.
Mol Microbiol ; 85(6): 1026-8, 2012 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-22812455

RESUMEN

LeuO is a quiescent genetic regulator present in many bacteria, which forms part of the H-NS regulon. LeuO in turn has been proposed to activate a subset of genes of the regulon by antagonizing H-NS. In the paper by Dillon et al., binding of LeuO to the S. Typhimurium genome was observed by ChIP-chip to some of the previously described LeuO-regulated genes, upon growth under stress conditions. However, studies at a higher LeuO concentration from a cloned inducible promoter rendered many more binding sites, pointing towards the importance of the abundance of the regulator in the cell, in a given moment. Binding of LeuO was observed not only to intergenic sequences, but in the majority of cases to intragenic sequences, and co-binding was observed with H-NS in many sites and with RNA polymerase to the majority of sites. The authors define a binding motif that allowed the detection of several other LeuO-regulated genes that were not detected by ChIP-chip, which were possibly missed because LeuO binds and bridges distal sites, in those instances. The observations reported open new questions regarding the mode of action for LeuO.


Asunto(s)
Regulación Bacteriana de la Expresión Génica , Salmonella typhimurium/genética , Factores de Transcripción/metabolismo
10.
BMC Microbiol ; 13: 264, 2013 Nov 21.
Artículo en Inglés | MEDLINE | ID: mdl-24262067

RESUMEN

BACKGROUND: Our observation that in the Mexican Salmonella Typhimurium population none of the ST19 and ST213 strains harbored both the Salmonella virulence plasmid (pSTV) and the prevalent IncA/C plasmid (pA/C) led us to hypothesize that restriction to horizontal transfer of these plasmids existed. We designed a conjugation scheme using ST213 strain YU39 as donor of the blaCMY-2 gene (conferring resistance to ceftriaxone; CRO) carried by pA/C, and two E. coli lab strains (DH5α and HB101) and two Typhimurium ST19 strains (SO1 and LT2) carrying pSTV as recipients. The aim of this study was to determine if the genetic background of the different recipient strains affected the transfer frequencies of pA/C. RESULTS: YU39 was able to transfer CRO resistance, via a novel conjugative mechanism, to all the recipient strains although at low frequencies (10-7 to 10-10). The presence of pSTV in the recipients had little effect on the conjugation frequency. The analysis of the transconjugants showed that three different phenomena were occurring associated to the transfer of blaCMY-2: 1) the co-integration of pA/C and pX1; 2) the transposition of the CMY region from pA/C to pX1; or 3) the rearrangement of pA/C. In addition, the co-lateral mobilization of a small (5 kb) ColE1-like plasmid was observed. The transconjugant plasmids involving pX1 re-arrangements (either via co-integration or ISEcp1-mediated transposition) obtained the capacity to conjugate at very high levels, similar to those found for pX1 (10-1). Two versions of the region containing blaCMY-2 were found to transpose to pX1: the large version was inserted into an intergenic region located where the "genetic load" operons are frequently inserted into pX1, while the short version was inserted into the stbDE operon involved in plasmid addiction system. This is the first study to report the acquisition of an extended spectrum cephalosporin (ESC)-resistance gene by an IncX1 plasmid. CONCLUSIONS: We showed that the transfer of the YU39 blaCMY-2 gene harbored on a non- conjugative pA/C requires the machinery of a highly conjugative pX1 plasmid. Our experiments demonstrate the complex interactions a single strain can exploit to contend with the challenge of horizontal transfer and antibiotic selective pressure.


Asunto(s)
Conjugación Genética , Transferencia de Gen Horizontal , Plásmidos , Salmonella typhimurium/enzimología , Salmonella typhimurium/genética , beta-Lactamasas/genética , Animales , ADN Bacteriano/química , ADN Bacteriano/genética , Escherichia coli/genética , Reordenamiento Génico , Humanos , México , Datos de Secuencia Molecular , Recombinación Genética , Infecciones por Salmonella/microbiología , Salmonelosis Animal/microbiología , Salmonella typhimurium/aislamiento & purificación , Análisis de Secuencia de ADN
11.
J Med Microbiol ; 71(1)2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-35006066

RESUMEN

Introduction. Salmonella enterica serovar Typhi (S. Typhi) is the etiological agent of typhoid fever. To establish an infection in the human host, this pathogen must survive the presence of bile salts in the gut and gallbladder.Hypothesis. S. Typhi uses multiple genetic elements to resist the presence of human bile.Aims. To determine the genetic elements that S. Typhi utilizes to tolerate the human bile salt sodium deoxycholate.Methodology. A collection of S. Typhi mutant strains was evaluated for their ability to growth in the presence of sodium deoxycholate and ox-bile. Additionally, transcriptomic and proteomic responses elicited by sodium deoxycholate on S. Typhi cultures were also analysed.Results. Multiple transcriptional factors and some of their dependent genes involved in central metabolism, as well as in cell envelope, are required for deoxycholate resistance.Conclusion. These findings suggest that metabolic adaptation to bile is focused on enhancing energy production to sustain synthesis of cell envelope components exposed to damage by bile salts.


Asunto(s)
Ácidos y Sales Biliares/química , Ácido Desoxicólico/química , Salmonella typhi , Bilis , Humanos , Proteómica , Salmonella typhi/metabolismo , Transcriptoma
12.
BMC Microbiol ; 11(1): 9, 2011 Jan 11.
Artículo en Inglés | MEDLINE | ID: mdl-21223599

RESUMEN

BACKGROUND: Salmonella Typhimurium ST213 was first detected in the Mexican Typhimurium population in 2001. It is associated with a multi-drug resistance phenotype and a plasmid-borne blaCMY-2 gene conferring resistance to extended-spectrum cephalosporins. The objective of the current study was to examine the association between the ST213 genotype and blaCMY-2 plasmids. RESULTS: The blaCMY-2 gene was carried by an IncA/C plasmid. ST213 strains lacking the blaCMY-2 gene carried a different IncA/C plasmid. PCR analysis of seven DNA regions distributed throughout the plasmids showed that these IncA/C plasmids were related, but the presence and absence of DNA stretches produced two divergent types I and II. A class 1 integron (dfrA12, orfF and aadA2) was detected in most of the type I plasmids. Type I contained all the plasmids carrying the blaCMY-2 gene and a subset of plasmids lacking blaCMY-2. Type II included all of the remaining blaCMY-2-negative plasmids. A sequence comparison of the seven DNA regions showed that both types were closely related to IncA/C plasmids found in Escherichia, Salmonella, Yersinia, Photobacterium, Vibrio and Aeromonas. Analysis of our Typhimurium strains showed that the region containing the blaCMY-2 gene is inserted between traA and traC as a single copy, like in the E. coli plasmid pAR060302. The floR allele was identical to that of Newport pSN254, suggesting a mosaic pattern of ancestry with plasmids from other Salmonella serovars and E. coli. Only one of the tested strains was able to conjugate the IncA/C plasmid at very low frequencies (10-7 to 10-9). The lack of conjugation ability of our IncA/C plasmids agrees with the clonal dissemination trend suggested by the chromosomal backgrounds and plasmid pattern associations. CONCLUSIONS: The ecological success of the newly emerging Typhimurium ST213 genotype in Mexico may be related to the carriage of IncA/C plasmids. We conclude that types I and II of IncA/C plasmids originated from a common ancestor and that the insertion and deletion of DNA stretches have shaped their evolutionary histories.


Asunto(s)
Resistencia a Múltiples Medicamentos/genética , Plásmidos/genética , Salmonella typhimurium/genética , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Southern Blotting , Genotipo , Reacción en Cadena de la Polimerasa , Salmonella typhimurium/efectos de los fármacos
13.
J Med Microbiol ; 70(9)2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34590996

RESUMEN

The S. Typhi leuO gene, which codes for the LysR-type transcriptional regulator LeuO, contains five forward promoters named P3, P1, P2, P5 and P4, and two reverse promoters, P6 and P7. The activity of the forward promoters was revealed by primer extension using gene reporter fusions in an S. Typhi hns lrp mutant strain. Likewise, the activity of the reverse promoters was revealed in an hns background. Derepression of the transcription of the chromosomal gene was confirmed by RT-PCR in the hns lrp mutant. The leuOP1 transcriptional reporter fusion, which contained only the major P1 promoter, had a lower expression in a relA spoT mutant strain, indicating that the steady-state levels of the (p)ppGpp alarmone positively regulate it. In contrast, the leuOP3, leuOP5P4, leuOP6 and leuOP7 transcriptional fusions were derepressed in the relA spoT background, indicating that the alarmone has a negative effect on their expression. Thus, the search for genetic regulators and environmental cues that would differentially derepress leuO gene expression by antagonizing the action of the H-NS and Lrp nucleoid-associated proteins, or that would fine-tune the expression of the various promoters, will further our understanding of the significance that multiple promoters have in the control of LeuO expression.


Asunto(s)
Proteínas Bacterianas/genética , Regiones Promotoras Genéticas , Salmonella typhi/genética , Factores de Transcripción/genética , Regiones Promotoras Genéticas/genética , Regiones Promotoras Genéticas/fisiología , ARN Bacteriano/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
14.
Front Microbiol ; 12: 657404, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33854491

RESUMEN

The CRISPR-Cas cluster is found in many prokaryotic genomes including those of the Enterobacteriaceae family. Salmonella enterica serovar Typhi (S. Typhi) harbors a Type I-E CRISPR-Cas locus composed of cas3, cse1, cse2, cas7, cas5, cas6e, cas1, cas2, and a CRISPR1 array. In this work, it was determined that, in the absence of cas5 or cas2, the amount of the OmpC porin decreased substantially, whereas in individual cse2, cas6e, cas1, or cas3 null mutants, the OmpF porin was not observed in an electrophoretic profile of outer membrane proteins. Furthermore, the LysR-type transcriptional regulator LeuO was unable to positively regulate the expression of the quiescent OmpS2 porin, in individual S. Typhi cse2, cas5, cas6e, cas1, cas2, and cas3 mutants. Remarkably, the expression of the master porin regulator OmpR was dependent on the Cse2, Cas5, Cas6e, Cas1, Cas2, and Cas3 proteins. Therefore, the data suggest that the CRISPR-Cas system acts hierarchically on OmpR to control the synthesis of outer membrane proteins in S. Typhi.

15.
BMC Microbiol ; 9: 131, 2009 Jul 03.
Artículo en Inglés | MEDLINE | ID: mdl-19573249

RESUMEN

BACKGROUND: Bacterial genomes are mosaic structures composed of genes present in every strain of the same species (core genome), and genes present in some but not all strains of a species (accessory genome). The aim of this study was to compare the genetic diversity of core and accessory genes of a Salmonella enterica subspecies enterica serovar Typhimurium (Typhimurium) population isolated from food-animal and human sources in four regions of Mexico. Multilocus sequence typing (MLST) and macrorestriction fingerprints by pulsed-field gel electrophoresis (PFGE) were used to address the core genetic variation, and genes involved in pathogenesis and antibiotic resistance were selected to evaluate the accessory genome. RESULTS: We found a low genetic diversity for both housekeeping and accessory genes. Sequence type 19 (ST19) was supported as the founder genotype of STs 213, 302 and 429. We found a temporal pattern in which the derived ST213 is replacing the founder ST19 in the four geographic regions analyzed and a geographic trend in the number of resistance determinants. The distribution of the accessory genes was not random among chromosomal genotypes. We detected strong associations among the different accessory genes and the multilocus chromosomal genotypes (STs). First, the Salmonella virulence plasmid (pSTV) was found mostly in ST19 isolates. Second, the plasmid-borne betalactamase cmy-2 was found only in ST213 isolates. Third, the most abundant integron, IP-1 (dfrA12, orfF and aadA2), was found only in ST213 isolates. Fourth, the Salmonella genomic island (SGI1) was found mainly in a subgroup of ST19 isolates carrying pSTV. The mapping of accessory genes and multilocus genotypes on the dendrogram derived from macrorestiction fingerprints allowed the establishment of genetic subgroups within the population. CONCLUSION: Despite the low levels of genetic diversity of core and accessory genes, the non-random distribution of the accessory genes across chromosomal backgrounds allowed us to discover genetic subgroups within the population. This study provides information about the importance of the accessory genome in generating genetic variability within a bacterial population.


Asunto(s)
Farmacorresistencia Bacteriana/genética , Variación Genética , Genoma Bacteriano , Salmonella typhimurium/genética , Técnicas de Tipificación Bacteriana , Cromosomas Bacterianos/genética , ADN Bacteriano/genética , Genes Bacterianos , Marcadores Genéticos , Genética de Población , Islas Genómicas , Genotipo , Integrones/genética , México , Pruebas de Sensibilidad Microbiana , Plásmidos/genética , Salmonella typhimurium/clasificación , Salmonella typhimurium/patogenicidad , Análisis de Secuencia de ADN , Virulencia/genética
16.
PLoS One ; 14(10): e0223975, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31626639

RESUMEN

The Salmonella enterica serovar Typhimurium sequence type 213 (ST213) emerged as a predominant genotype in Mexico. It is characterized by harboring multidrug resistance (MDR) IncC plasmids (previously IncA/C) and the lack of the Salmonella virulence plasmid (pSTV). Here we show that the D6-like plasmid prophage is present in most of the ST213 strains. We used the reported nucleotide sequence of YU39 plasmid (pYU39_89) to design a PCR typing scheme for the D6-like plasmid prophages, and determined the complete nucleotide sequences for the D6-like prophages of three additional ST213 strains (YU07-18, SL26 and SO21). Two prophage variants were described: i) a complete prophage, containing homologous sequences for most of the genetic modules described in P1 and D6 phages, which most likely allow for the lytic and lysogenic lifestyles; and ii) an incomplete prophage, lacking a 15 kb region containing morphogenesis genes, suggesting that it is defective. The tail fiber gene inversion region was the most divergent one between D6 and pYU39_89 genomes, suggesting the production of a distinct set of tail fibers, which could be involved in host range preferences. A glutaminyl-tRNA synthetase gene (glnS), which could be involved in providing host cell increased fitness or plasmid maintenance functions, was found in all D6-like genomes. Population level analysis revealed a biogeographic pattern of distribution of these plasmid-phages and specific associations with variants of MDR IncC plasmids. Statistically significant associations were found between the two prophage variants (p75 or p89), the type of IncC plasmids (I or II) and geographic isolation regions (Sonora, San Luis Potosí, Michoacán and Yucatán). This work integrates results from molecular typing, genomics and epidemiology to provide a broad overview for the evolution of an emergent Salmonella genotype.


Asunto(s)
Plásmidos/metabolismo , Profagos/fisiología , Salmonella typhimurium/patogenicidad , Aminoacil-ARNt Sintetasas/genética , Evolución Molecular , Genoma Viral , Genómica/métodos , Genotipo , Plásmidos/genética , Reacción en Cadena de la Polimerasa , Profagos/genética , Profagos/aislamiento & purificación , Salmonella typhimurium/genética , Salmonella typhimurium/virología , Proteínas Virales/genética , Virulencia/genética
17.
Pathog Dis ; 76(1)2018 02 01.
Artículo en Inglés | MEDLINE | ID: mdl-29325038

RESUMEN

In nature, microorganisms are constantly exposed to multiple viral infections and thus have developed many strategies to survive phage attack and invasion by foreign DNA. One of such strategies is the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) bacterial immunological system. This defense mechanism is widespread in prokaryotes including several families such as Enterobacteriaceae. Much knowledge about the CRISPR-Cas system has been generated, including its biological functions, transcriptional regulation, distribution, utility as a molecular marker and as a tool for specific genome editing. This review focuses on these aspects and describes the state of the art of the CRISPR-Cas system in the Enterobacteriaceae bacterial family.


Asunto(s)
Sistemas CRISPR-Cas , Enterobacteriaceae/enzimología , Enterobacteriaceae/genética , Regulación de la Expresión Génica , Variación Genética
18.
Pathog Dis ; 2017 Jun 22.
Artículo en Inglés | MEDLINE | ID: mdl-28645187

RESUMEN

A current view on the role of the Salmonella virulence plasmid in the pathogenesis of animal and human hosts is discussed; including the possible relevance in secondary ecological niches. Various strategies towards further studies in this respect are proposed within the One Health Concept.

19.
Int J Hyg Environ Health ; 209(5): 461-70, 2006 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-16762593

RESUMEN

Fecal pollution of settled dust samples from indoor and outdoor urban environments, was measured and characterized by the presence of fecal coliforms (FC), and by the characterization of Escherichia coli virulence genes, adherence and antibiotic resistance traits as markers. There were more FC indoors than outdoors (mean values 1089 and 435MPN/g). Among indoor samples, there were more FC in houses with carpets and/or pets. Using a PCR-based assay for six enteropathogenicity genes (belonging to the EAEC, EHEC and EPEC pathotypes) on randomly selected E. coli isolates, there was no significant difference between isolates from indoors and outdoors (60% and 72% positive to at least one gene). The serotypes commonly associated with pathogenic strains, such as O86 and O28, were found in the indoor isolates; whereas O4, O66 and O9 were found amongst outdoor isolates. However, there were significantly more outdoor isolates resistant to at least one antibiotic (73% vs. 45% from indoors) among the strains positive for virulence factors, and outdoor isolates were more commonly multiresistant. There was no relationship between the presence of virulence genes and resistance traits. These results indicate that outdoor fecal bacteria were more likely from human sources, and those found indoors were related to pets and maintained in carpets. This study illustrates the risk posed by fecal bacteria from human sources, usually bearing virulence and resistance traits. Furthermore, the high prevalence of strains carrying genes associated to EAEC or EHEC pathotypes, in both indoor and outdoor environments, adds to the health risk.


Asunto(s)
Polvo , Contaminantes Ambientales , Escherichia coli , Farmacorresistencia Bacteriana , Polvo/análisis , Contaminantes Ambientales/análisis , Escherichia coli/clasificación , Escherichia coli/efectos de los fármacos , Escherichia coli/patogenicidad , Humanos , México , Características de la Residencia , Población Urbana , Virulencia
20.
Genome Announc ; 4(2)2016 Apr 21.
Artículo en Inglés | MEDLINE | ID: mdl-27103717

RESUMEN

The complete genome of ITALIC! Salmonella entericaserovar Typhimurium strain SO3 (sequence type 302), isolated from a fatal meningitis infection in Mexico, was determined using PacBio technology. The chromosome hosts six complete prophages and is predicted to harbor 51 genomic islands, including 13 pathogenicity islands (SPIs). It carries the ITALIC! Salmonellavirulence plasmid (pSTV).

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