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1.
J Bacteriol ; 206(1): e0030923, 2024 01 25.
Artículo en Inglés | MEDLINE | ID: mdl-38179929

RESUMEN

In addition to its role in genome protection, DNA methylation can regulate gene expression. In this study, we characterized the impact of acidity, phase variation, and the ArsRS TCS on the expression of the Type I m6A DNA methyltransferase HsdM1 (HP0463) of Helicobacter pylori 26695 and their subsequent effects on the methylome. Transcription of hsdM1 increases at least fourfold in the absence of the sensory histidine kinase ArsS, the major acid-sensing protein of H. pylori. hsdM1 exists in the phase-variable operon hsdR1-hsdM1. Phase-locking hsdR1 (HP0464), the restriction endonuclease gene, has significant impacts on the transcription of hsdM1. To determine the impacts of methyltransferase transcription patterns on the methylome, we conducted methylome sequencing on samples cultured at pH 7 or pH 5. We found differentially methylated motifs between these growth conditions and that deletions of arsS and/or hsdM1 interfere with the epigenetic acid response. Deletion of arsS leads to altered activity of HsdM1 and multiple other methyltransferases under both pH conditions indicating that the ArsRS TCS, in addition to direct effects on regulon transcription during acid acclimation, may also indirectly impact gene expression via regulation of the methylome. We determined the target motif of HsdM1 (HP0463) to be the complementary bipartite sequence pair 5'-TCAm6AVN6TGY-3' and 3'-AGTN6GAm6ACA-5'. This complex regulation of DNA methyltransferases, and thus differential methylation patterns, may have implications for the decades-long persistent infection by H. pylori. IMPORTANCE This study expands the possibilities for complex, epigenomic regulation in Helicobacter pylori. We demonstrate that the H. pylori methylome is plastic and acid sensitive via the two-component system ArsRS and the DNA methyltransferase HsdM1. The control of a methyltransferase by ArsRS may allow for a layered response to changing acidity. Likely, an early response whereby ArsR~P affects regulon expression, including the methyltransferase hsdM1. Then, a somewhat later effect as the altered methylome, due to altered HsdM1 expression, subsequently alters the expression of other genes involved in acclimation. The intermediate methylation of certain motifs supports the hypothesis that methyltransferases play a regulatory role. Untangling this additional web of regulation could play a key role in understanding H. pylori colonization and persistence.


Asunto(s)
Helicobacter pylori , Helicobacter pylori/genética , Helicobacter pylori/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Epigenoma , Metiltransferasas/genética , ADN/metabolismo , Regulación Bacteriana de la Expresión Génica , Metilación de ADN
2.
Infect Immun ; 89(4)2021 03 17.
Artículo en Inglés | MEDLINE | ID: mdl-33526561

RESUMEN

Helicobacter pylori encounters a wide range of pH within the human stomach. In a comparison of H. pylori cultured in vitro under neutral or acidic conditions, about 15% of genes are differentially expressed, and corresponding changes are detectable for many of the encoded proteins. The ArsRS two-component system (TCS), comprised of the sensor kinase ArsS and its cognate response regulator ArsR, has an important role in mediating pH-responsive changes in H. pylori gene expression. In this study, we sought to delineate the pH-responsive ArsRS regulon and further define the role of ArsR in pH-responsive gene expression. We compared H. pylori strains containing an intact ArsRS system with an arsS null mutant or strains containing site-specific mutations of a conserved aspartate residue (D52) in ArsR, which is phosphorylated in response to signals relayed by the cognate sensor kinase ArsS. We identified 178 genes that were pH-responsive in strains containing an intact ArsRS system but not in ΔarsS or arsR mutants. These constituents of the pH-responsive ArsRS regulon include genes involved in acid acclimatization (ureAB, amidases), oxidative stress responses (katA, sodB), transcriptional regulation related to iron or nickel homeostasis (fur, nikR), and genes encoding outer membrane proteins (including sabA, alpA, alpB, hopD [labA], and horA). When comparing H. pylori strains containing an intact ArsRS TCS with arsRS mutants, each cultured at neutral pH, relatively few genes are differentially expressed. Collectively, these data suggest that ArsRS-mediated gene regulation has an important role in H. pylori adaptation to changing pH conditions.


Asunto(s)
Regulación Bacteriana de la Expresión Génica , Helicobacter pylori/fisiología , Concentración de Iones de Hidrógeno , Elementos de Respuesta , Transactivadores/metabolismo , Biología Computacional/métodos , Perfilación de la Expresión Génica , Humanos , Mutación , Proteoma , Proteómica/métodos , Transcripción Genética
3.
J Infect Dis ; 214(4): 644-8, 2016 08 15.
Artículo en Inglés | MEDLINE | ID: mdl-27190191

RESUMEN

Helicobacter pylori is the strongest risk factor for gastric adenocarcinoma, which develops within a hypochlorhydric environment. We sequentially isolated H. pylori (strain J99) from a patient who developed corpus-predominant gastritis and hypochlorhydia over a 6-year interval. Archival J99 survived significantly better under acidic conditions than recent J99 strains. H. pylori arsRS encodes a 2-component system critical for stress responses; recent J99 isolates harbored 2 nonsynonymous arsS mutations, and arsS inactivation abolished acid survival. In vivo, acid-resistant archival, but not recent J99, successfully colonized high-acid-secreting rodents. Thus, genetic evolution of arsS may influence progression to hypochlorhydia and gastric cancer.


Asunto(s)
Aclorhidria/microbiología , Evolución Molecular , Gastritis/microbiología , Infecciones por Helicobacter/microbiología , Helicobacter pylori/enzimología , Helicobacter pylori/genética , Histidina Quinasa/genética , Ácidos/toxicidad , Animales , Proteínas Bacterianas/genética , Gastritis/complicaciones , Gerbillinae , Helicobacter pylori/aislamiento & purificación , Helicobacter pylori/fisiología , Humanos , Masculino , Ratones Endogámicos C57BL , Viabilidad Microbiana/efectos de los fármacos , Mutación Missense
4.
J Bacteriol ; 196(19): 3421-9, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-25022855

RESUMEN

The pathogenesis of diseases elicited by the gastric pathogen Helicobacter pylori is partially determined by the effectiveness of adaptation to the variably acidic environment of the host stomach. Adaptation includes appropriate adherence to the gastric epithelium via outer membrane protein adhesins such as SabA. The expression of sabA is subject to regulation via phase variation in the promoter and coding regions as well as repression by the two-component system ArsRS. In this study, we investigated the role of a homopolymeric thymine [poly(T)] tract -50 to -33 relative to the sabA transcriptional start site in H. pylori strain J99. We quantified sabA expression in H. pylori J99 by quantitative reverse transcription-PCR (RT-PCR), demonstrating significant changes in sabA expression associated with experimental manipulations of poly(T) tract length. Mimicking the length increase of this tract by adding adenines instead of thymines had similar effects, while the addition of other nucleotides failed to affect sabA expression in the same manner. We hypothesize that modification of the poly(T) tract changes DNA topology, affecting regulatory protein interaction(s) or RNA polymerase binding efficiency. Additionally, we characterized the interaction between the sabA promoter region and ArsR, a response regulator affecting sabA expression. Using recombinant ArsR in electrophoretic mobility shift assays (EMSA), we localized binding to a sequence with partial dyad symmetry -20 and +38 relative to the sabA +1 site. The control of sabA expression by both ArsRS and phase variation at two distinct repeat regions suggests the control of sabA expression is both complex and vital to H. pylori infection.


Asunto(s)
Adhesinas Bacterianas/genética , Helicobacter pylori/genética , Regiones Promotoras Genéticas , Secuencias Repetitivas de Ácidos Nucleicos , Transcripción Genética , Adhesinas Bacterianas/metabolismo , Secuencia de Bases , Regulación Bacteriana de la Expresión Génica , Helicobacter pylori/metabolismo , Datos de Secuencia Molecular
5.
Microbiol Spectr ; 11(1): e0463322, 2023 02 14.
Artículo en Inglés | MEDLINE | ID: mdl-36625670

RESUMEN

Helicobacter pylori colonizes the human gastric mucosa and causes various gastroduodenal diseases, including peptic ulceration and gastric cancer. Colonization requires the actions of two-component systems (TCSs) to sense and respond to changes in the host environment. In this study, we evaluated gene regulation mediated by the CrdRS TCS. Few studies have evaluated this TCS, leaving the signal(s) yet to be exhaustively determined and a need for a more complete regulon to be delineated. We performed RNA sequencing (RNA-Seq) on three isogenic H. pylori 26695 mutants: a control, a mutant with deletion of the sensory histidine kinase, ΔcrdS, and a mutant with deletion of the response regulator, ΔcrdR. Comparison of the RNA-Seq results from these mutants established a 40-gene regulon putatively controlled by the CrdRS TCS. Quantitative reverse transcriptase PCR (RT-qPCR) was used to validate 7 of 11 putative regulon members selected for analysis. We further investigated 6 confirmed CrdRS regulon genes by using phospho-incompetent H. pylori 26695 CrdR D53A and CrdS H173A mutants. These experiments further confirmed the role of CrdRS in regulation of urease, acetone carboxylase, hofD, and HP1440. Expression of these CrdRS regulon genes was also evaluated under 10 µM nitric oxide (NO) conditions. This revealed that ureA, acxA, hofD, and HP1440 expression is affected by NO in a CrdRS-dependent manner. Importantly, three of these genes (ureA, acxA, and hofD) are known to play important roles in H. pylori colonization of the stomach. IMPORTANCE The molecular strategies used by Helicobacter pylori to colonize and persist in the harsh environment of the human stomach are a critical area of study. Our study identified several genes in this gastric pathogen, including ureA, a gene encoding a protein essential to the survival of H. pylori, that are regulated via the CrdRS two-component system (TCS) in response to nitric oxide (NO). NO is a product of the innate immune system of the human host. The identification of these genes whose expression is regulated by this molecule may give insights to novel therapeutics. Two genes (ureA and acxA) determined in this study to be regulated by NO via CrdRS have been previously determined to be regulated by other TCSs, indicating that the expression of these genes may be of critical importance to H. pylori.


Asunto(s)
Infecciones por Helicobacter , Helicobacter pylori , Humanos , Ureasa/genética , Ureasa/metabolismo , Helicobacter pylori/genética , Helicobacter pylori/metabolismo , Óxido Nítrico , Proteínas Bacterianas/metabolismo , Infecciones por Helicobacter/genética
6.
Microb Pathog ; 53(5-6): 227-33, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22940419

RESUMEN

Phase variation is frequently utilized by bacterial species to affect gene expression such that phenotypic variants are maintained within populations, ensuring survival as environmental or host conditions change. Unusual among Helicobacter pylori phase variable or contingency genes is arsS, encoding a sensory histidine kinase involved in the acid acclimation of the organism. The presence of a 3' homopolymeric cytosine tract of variable length in arsS among Helicobacter pylori strains allows for the expression of various functional ArsS isoforms, differing in carboxy-terminal protein domains. In this study, we analyzed this 3'arsS region via amplified fragment length polymorphism (AFLP) and sequencing analyses for H. pylori populations from 3 different gastric sites of 12 patients. Our data indicate the presence of multiple arsS alleles within each population of H. pylori derived from the gastric antrum, cardia, or corpus of these patients. We also show that H. pylori, derived from the same anatomical site and patient, are predicted to express multiple ArsS isoforms in each population investigated. Furthermore, we identify a polymorphic deletion within arsS that generates another alternate ArsS C-terminal end. These findings suggest that four C-terminal variations of ArsS adds to the complexity of the ArsRS acid adaptation mechanism as a whole and may influence the ability of H. pylori to persist in the gastric niche for decades.


Asunto(s)
Mucosa Gástrica/microbiología , Helicobacter pylori/enzimología , Helicobacter pylori/genética , Polimorfismo Genético , Proteínas Quinasas/genética , Anciano , Alelos , Análisis del Polimorfismo de Longitud de Fragmentos Amplificados , Citidina/genética , ADN Bacteriano/química , ADN Bacteriano/genética , Femenino , Genotipo , Helicobacter pylori/aislamiento & purificación , Histidina Quinasa , Humanos , Masculino , Persona de Mediana Edad , Análisis de Secuencia de ADN , Eliminación de Secuencia
7.
Pathog Dis ; 75(8)2017 11 30.
Artículo en Inglés | MEDLINE | ID: mdl-29040466

RESUMEN

OipA is a phase-variable virulence factor of Helicobacter pylori. Mutations in oipA to turn the gene phase on in a cag pathogenicity island (PAI)-negative strain of H. pylori (J68) or phase off in a cag PAI-positive strain (26695) demonstrated that phase on oipA alleles in both strains had both increased oipA mRNA and human gastric adenocarcinoma (AGS) cell adherence compared to isogenic oipA phase off mutants. An oipA phase off mutant of H. pylori 26695 demonstrated decreased IL-8 secretion by AGS cells and failure to translocate the cag PAI effector CagA. Increased attachment by OipA expressing cag PAI-negative H. pylori J68 failed to alter secreted IL-8 levels. Thus, OipA is necessary but not sufficient for the induction of IL-8; however, it is necessary for translocation of the oncoprotein CagA. Perhaps the nearly invariant phase on status of oipA alleles among cag PAI-positive H. pylori isolates relates to the role of this outer membrane protein in effective translocation of CagA. oipA mRNA comparisons between AGS cell-adherent and non-adherent H. pylori 26695 revealed significantly greater levels in the adherent cells. This may allow H. pylori to adapt to conditions of host cell contact by altering expression of this virulence factor.


Asunto(s)
Antígenos Bacterianos/metabolismo , Proteínas de la Membrana Bacteriana Externa/metabolismo , Proteínas Bacterianas/metabolismo , Islas Genómicas/genética , Helicobacter pylori/patogenicidad , Interleucina-8/metabolismo , Sistemas de Secreción Tipo IV/genética , Análisis del Polimorfismo de Longitud de Fragmentos Amplificados , Antígenos Bacterianos/genética , Adhesión Bacteriana/fisiología , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas Bacterianas/genética , Línea Celular Tumoral , Infecciones por Helicobacter/microbiología , Infecciones por Helicobacter/patología , Helicobacter pylori/genética , Interacciones Huésped-Patógeno , Humanos , Interleucina-8/inmunología , Transporte de Proteínas/fisiología , ARN Mensajero/biosíntesis , ARN Mensajero/genética
8.
J Med Microbiol ; 66(6): 798-807, 2017 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-28598306

RESUMEN

PURPOSE: We investigated the transcription of adhesin-encoding genes sabA, hopZ and labA in Helicobacter pylori strain J99. Each possesses a repeating homopolymeric nucleotide tract within their promoter regions, and sabA and hopZ possess repeats within their 5' coding regions. METHODOLOGY: We altered the repeat lengths associated with the adhesin genes and quantified mRNA levels by real-time quantitative PCR. Using adherence to AGS cells and IL-8 assays, we examined the effects of altered transcript levels. We assessed the role of ArsRS in transcription using an arsS null mutant and by examining ArsR binding to promoter regions via electrophoretic mobility shift assays. RESULTS: Extensions or truncations of promoter region repeats in hopZ and labA increased transcript levels, mirroring results shown by our lab and others for mutations in the sabA promoter. Altered lengths of the poly-cytosine thymine tract within the 5' coding region of sabA demonstrated that switching from phase-off to phase-on significantly increased mRNA levels. However, mutations in the poly-thymine tract of sabA, which increased mRNA levels, do not behave synergistically with phase-on mutations. Phase-on mutations of sabA resulted in increased H. pylori adherence to AGS cells, but only a modest effect on IL-8. hopZ and labA, and sabA paralogue sabB, transcript levels were increased in an arsS mutant and ArsR bound the promoter regions for each of these genes in vitro. CONCLUSION: This work highlights the complex nature of adhesin regulation, its impact on H. pylori attachment and the pervasive role of ArsRS in adhesin expression. Such regulation may help facilitate the decades-long persistence of infection.


Asunto(s)
Adhesinas Bacterianas/genética , Proteínas Bacterianas/genética , Regulación Bacteriana de la Expresión Génica , Sistemas de Lectura Abierta , Regiones Promotoras Genéticas , Adhesión Bacteriana/genética , Proteínas de la Membrana Bacteriana Externa/genética , Línea Celular , Helicobacter pylori/genética , Histidina Quinasa/genética , Histidina Quinasa/metabolismo , Humanos , Interleucina-8 , Mutación , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Transcripción Genética
9.
PLoS One ; 12(11): e0188804, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-29186206

RESUMEN

Helicobacter pylori is a genetically diverse bacterial species that colonizes the stomach in about half of the human population. Most persons colonized by H. pylori remain asymptomatic, but the presence of this organism is a risk factor for gastric cancer. Multiple populations and subpopulations of H. pylori with distinct geographic distributions are recognized. Genetic differences among these populations might be a factor underlying geographic variation in gastric cancer incidence. Relatively little is known about the genomic features of African H. pylori strains compared to other populations of strains. In this study, we first analyzed the genomes of H. pylori strains from seven globally distributed populations or subpopulations and identified encoded proteins that exhibited the highest levels of sequence divergence. These included secreted proteins, an LPS glycosyltransferase, fucosyltransferases, proteins involved in molybdopterin biosynthesis, and Clp protease adaptor (ClpS). Among proteins encoded by the cag pathogenicity island, CagA and CagQ exhibited the highest levels of sequence diversity. We then identified proteins in strains of Western African origin (classified as hspWAfrica by MLST analysis) with sequences that were highly divergent compared to those in other populations of strains. These included ATP-dependent Clp protease, ClpS, and proteins of unknown function. Three of the divergent proteins sequences identified in West African strains were characterized by distinct insertions or deletions up to 8 amino acids in length. These polymorphisms in rapidly evolving proteins represent robust genetic signatures for H. pylori strains of West African origin.


Asunto(s)
Helicobacter pylori/genética , África Occidental , Secuencia de Aminoácidos , Proteínas Bacterianas/química , Genes Bacterianos , Homología de Secuencia de Aminoácido
10.
Genome Announc ; 5(49)2017 Dec 07.
Artículo en Inglés | MEDLINE | ID: mdl-29217789

RESUMEN

We report the complete genome sequences of 19 cluster CA bacteriophages isolated from environmental samples using Rhodococcus erythropolis as a host. All of the phages are Siphoviridae, have similar genome lengths (46,314 to 46,985 bp) and G+C contents (58.5 to 58.8%), and share nucleotide sequence similarity.

12.
Front Microbiol ; 6: 558, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26124751

RESUMEN

Helicobacter pylori NikR (HpNikR) is a nickel dependent transcription factor that directly regulates a number of genes in this important gastric pathogen. One key gene that is regulated by HpNikR is ureA, which encodes for the urease enzyme. In vitro DNA binding studies of HpNikR with the ureA promoter (PureA ) previously identified a recognition site that is required for high affinity protein/DNA binding. As a means to determine the in vivo significance of this recognition site and to identify the key DNA sequence determinants required for ureA transcription, herein, we have translated these in vitro results to analysis directly within H. pylori. Using a series of GFP reporter constructs in which the PureA DNA target was altered, in combination with mutant H. pylori strains deficient in key regulatory proteins, we confirmed the importance of the previously identified HpNikR recognition sequence for HpNikR-dependent ureA transcription. Moreover, we identified a second factor, the HpArsRS two-component system that was required for maximum transcription of ureA. While HpArsRS is known to regulate ureA in response to acid shock, it was previously thought to function independently of HpNikR and to have no role at neutral pH. However, our qPCR analysis of ureA expression in wildtype, ΔnikR and ΔarsS single mutants as well as a ΔarsS/nikR double mutant strain background showed reduced basal level expression of ureA when arsS was absent. Additionally, we determined that both HpNikR and HpArsRS were necessary for maximal expression of ureA under nickel, low pH and combined nickel and low pH stresses. In vitro studies of HpArsR-P with the PureA DNA target using florescence anisotropy confirmed a direct protein/DNA binding interaction. Together, these data support a model in which HpArsRS and HpNikR cooperatively interact to regulate ureA transcription under various environmental conditions. This is the first time that direct "cross-talk" between HpArsRS and HpNikR at neutral pH has been demonstrated.

13.
PLoS One ; 6(10): e26750, 2011.
Artículo en Inglés | MEDLINE | ID: mdl-22053209

RESUMEN

Five newly isolated mycobacteriophages--Angelica, CrimD, Adephagia, Anaya, and Pixie--have similar genomic architectures to mycobacteriophage TM4, a previously characterized phage that is widely used in mycobacterial genetics. The nucleotide sequence similarities warrant grouping these into Cluster K, with subdivision into three subclusters: K1, K2, and K3. Although the overall genome architectures of these phages are similar, TM4 appears to have lost at least two segments of its genome, a central region containing the integration apparatus, and a segment at the right end. This suggests that TM4 is a recent derivative of a temperate parent, resolving a long-standing conundrum about its biology, in that it was reportedly recovered from a lysogenic strain of Mycobacterium avium, but it is not capable of forming lysogens in any mycobacterial host. Like TM4, all of the Cluster K phages infect both fast- and slow-growing mycobacteria, and all of them--with the exception of TM4--form stable lysogens in both Mycobacterium smegmatis and Mycobacterium tuberculosis; immunity assays show that all five of these phages share the same immune specificity. TM4 infects these lysogens suggesting that it was either derived from a heteroimmune temperate parent or that it has acquired a virulent phenotype. We have also characterized a widely-used conditionally replicating derivative of TM4 and identified mutations conferring the temperature-sensitive phenotype. All of the Cluster K phages contain a series of well conserved 13 bp repeats associated with the translation initiation sites of a subset of the genes; approximately one half of these contain an additional sequence feature composed of imperfectly conserved 17 bp inverted repeats separated by a variable spacer. The K1 phages integrate into the host tmRNA and the Cluster K phages represent potential new tools for the genetics of M. tuberculosis and related species.


Asunto(s)
Evolución Molecular , Micobacteriófagos/genética , Sitios de Ligazón Microbiológica , Secuencia de Bases , Mapeo Cromosómico , Análisis por Conglomerados , Secuencia Conservada/genética , Eliminación de Gen , Genoma Viral/genética , Anotación de Secuencia Molecular , Datos de Secuencia Molecular , Familia de Multigenes/genética , Mutación/genética , Micobacteriófagos/crecimiento & desarrollo , Micobacteriófagos/aislamiento & purificación , Micobacteriófagos/ultraestructura , Análisis de Secuencia de ADN , Temperatura , Proteínas Virales/genética , Virión/genética , Virión/ultraestructura , Integración Viral/genética , Replicación Viral/fisiología
14.
Microbiology (Reading) ; 154(Pt 8): 2231-2240, 2008 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-18667556

RESUMEN

Adaptation to the acidic microenvironment, and adherence to mucosal epithelium, are essential for persistent colonization of the human stomach by Helicobacter pylori. The expression of SabA, an adhesin implicated in the ability of H. pylori to adhere to the host gastric epithelium, can be modulated by phase variation via slipped-strand mispairing in repetitive nucleotide tracts located in both the promoter region and the coding region. This study demonstrates the occurrence of phase variation at the sabA locus within individual strains of H. pylori, and among multiple isolates from a single patient. In addition, transcription of sabA is repressed by the acid-responsive ArsRS two-component signal transduction system in vitro. Our results demonstrate that isogenic inactivation of the arsS (jhp0151/HP0165) histidine kinase locus results in a 10-fold SabA-dependent increase in adherence to gastric epithelial cells in strain J99 (contains an in-frame sabA allele), but not in strain 26695 (out-of-frame sabA allele). The combination of transcriptional regulation of the sabA locus by the ArsRS two-component signal-transduction system and the generation of subpopulations harbouring alternate sabA alleles by slipped-strand mispairing during chromosomal replication could permit H. pylori to rapidly adapt to varying microenvironments or host immune responses. As a pathogen with a paucity of regulatory proteins, this dual regulation indicates that SabA expression is a tightly regulated process in H. pylori infection.


Asunto(s)
Adhesinas Bacterianas/metabolismo , Proteínas Bacterianas/metabolismo , Regulación Bacteriana de la Expresión Génica , Infecciones por Helicobacter/microbiología , Helicobacter pylori/metabolismo , Proteínas Quinasas/metabolismo , Transducción de Señal , Adhesinas Bacterianas/genética , Adhesión Bacteriana , Proteínas Bacterianas/genética , Línea Celular Tumoral , Mucosa Gástrica/microbiología , Helicobacter pylori/genética , Histidina Quinasa , Humanos , Fenotipo , Proteínas Quinasas/genética
15.
Infect Immun ; 73(6): 3794-8, 2005 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-15908415

RESUMEN

In an analysis of Helicobacter pylori genomic DNA by macroarray methodology, genomic DNA from a panel of cag pathogenicity island (PAI)-negative H. pylori clinical isolates failed to hybridize with 27 genes located outside the cag PAI in a cag PAI-positive reference strain. PCR analyses confirmed that HP0217 (encoding a lipopolysaccharide biosynthetic protein) and HP1079 (encoding a protein of unknown function) were present significantly more frequently in cagA-positive strains than in cagA-negative strains. A low G+C content of these two genes suggests they were acquired by horizontal transfer events.


Asunto(s)
Antígenos Bacterianos/genética , Proteínas Bacterianas/genética , Genoma Bacteriano , Islas Genómicas , Helicobacter pylori/genética , Biomarcadores , Transferencia de Gen Horizontal , Helicobacter pylori/patogenicidad , Reacción en Cadena de la Polimerasa
16.
Infect Immun ; 72(9): 5506-10, 2004 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-15322055

RESUMEN

LuxS plays a role in the synthesis of an extracellular signaling molecule, autoinducer 2 (AI-2). To analyze a possible role of AI-2 in regulating Helicobacter pylori gene expression, we constructed a panel of transcriptional reporter strains. We show that the expression of H. pylori flaA is growth phase dependent and that flaA transcription increases in association with increased culture density. Mutating the luxS gene eliminates growth-phase-dependent control of flaA, and this growth phase dependence is restored when the luxS mutant strain is complemented with the wild-type luxS gene.


Asunto(s)
Proteínas Bacterianas/metabolismo , Flagelina/metabolismo , Regulación Bacteriana de la Expresión Génica , Helicobacter pylori/crecimiento & desarrollo , Homoserina/análogos & derivados , Proteínas Bacterianas/genética , Liasas de Carbono-Azufre , Medios de Cultivo , Flagelina/genética , Helicobacter pylori/genética , Helicobacter pylori/metabolismo , Homoserina/genética , Homoserina/metabolismo , Lactonas/metabolismo , Mutación
17.
J Bacteriol ; 184(16): 4630-5, 2002 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-12142435

RESUMEN

To identify putative members of a regulon controlled by the H. pylori sensory histidine kinase HP0164 (HK0164), we constructed HK0164 null mutant H. pylori strains and analyzed bacterial gene transcription using DNA arrays. Seven genes were differentially expressed in multiple HK0164 mutant strains compared to their expression in control strains. Strain-dependent variations in differential expression were also detected. These results indicate that the signal transduction circuit utilizing HK0164 controls the transcription of at least seven genes in H. pylori.


Asunto(s)
Genoma Bacteriano , Helicobacter pylori/enzimología , Helicobacter pylori/genética , Proteínas Quinasas/genética , Regulón/genética , Regulación Bacteriana de la Expresión Génica/fisiología , Histidina Quinasa , Mutación/fisiología , Análisis de Secuencia por Matrices de Oligonucleótidos , Transducción de Señal/genética , Transcripción Genética/genética
18.
Mol Microbiol ; 48(5): 1225-39, 2003 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-12787351

RESUMEN

To identify Helicobacter pylori genes with expression that is enhanced under low pH conditions, we used subtractive hybridization methodology. We identified 28 acid-induced genes, of which 18 have known or putative functions. Six pairs of genes were co-transcribed. Primer extension analysis identified single or multiple transcriptional start points (tsp) for 14 of the 22 loci. Sequence analysis of the -10 regions upstream of the tsps revealed consensus motifs for multiple RNA polymerase sigma factors present in H. pylori (sigma80, sigma54 and sigma28). No sequences resembling the -35 Escherichia coli consensus sequence (TTGACA) were present upstream of any of the genes. Both increased gene transcription and decreased mRNA decay contribute to the observed increase in H. pylori transcript abundance at acid pH. These studies document the complex response of H. pylori to environmental pH changes, and provide insight into mechanisms used for intragastric survival.


Asunto(s)
Proteínas Bacterianas/metabolismo , Helicobacter pylori/fisiología , Regiones Promotoras Genéticas/genética , Regulación hacia Arriba , Proteínas Bacterianas/genética , Secuencia de Bases , Regulación Bacteriana de la Expresión Génica , Helicobacter pylori/genética , Helicobacter pylori/metabolismo , Humanos , Concentración de Iones de Hidrógeno , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transcripción Genética
19.
J Clin Microbiol ; 42(12): 5658-63, 2004 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-15583296

RESUMEN

Helicobacter pylori is a genetically diverse bacterial species that chronically infects human stomachs and sometimes causes severe gastroduodenal disease. Studies of polymorphic DNA sequences can suggest geographic origins of individual strains. Here, we describe a 180-bp insertion (ins180), which is just after the translation stop of a gene of unknown function, near the promoter of jhp0152-jhp0151 two-component signal transduction genes in strain J99, and absent from this site in strain 26695. This ins180 insertion was found in 9 of 9 Gambian (West African), 9 of 20 (45%) South African, and 9 of 40 (23%) Spanish strains but in only 2 of 20 (10%) North American strains and none of 20 Lithuanian, 20 Indian, and 20 Japanese strains. Four South African isolates that lacked ins180 and that belonged to an unusual outlier group contained a 480-bp insertion at this site (ins480), whereas none of 181 other strains screened contained ins480. In further tests 56% (10 of 18) of strains from African Americans but only 17% (3 of 18) of strains from Caucasian Americans carried ins180 (P < 0.05). Thus, the H. pylori strains of modern African Americans seem to retain traces of African roots, despite the multiple generations since their ancestors were taken from West Africa. Fragmentary ins180-like sequences were found at numerous sites in H. pylori genomes, always between genes. Such sequences might affect regulation of transcription and could facilitate genome rearrangement by homologous recombination. Apparent differences between African-American and Caucasian-American H. pylori gene pools may bear on our understanding of H. pylori transmission and disease outcome.


Asunto(s)
Emparejamiento Base/genética , Población Negra , Negro o Afroamericano , Elementos Transponibles de ADN/genética , Infecciones por Helicobacter/etnología , Helicobacter pylori/genética , Secuencia de Bases , Gambia , Infecciones por Helicobacter/genética , Infecciones por Helicobacter/microbiología , Humanos , Lituania , Datos de Secuencia Molecular , Análisis de Secuencia de ADN , Sudáfrica , Población Blanca
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