RESUMEN
Isotopic distributions of fragments from fission of the neutron-deficient ^{178}Hg nuclide are reported. This experimental observable is obtained for the first time in the region around lead using an innovative approach based on inverse kinematics and the coincidence between the large acceptance magnetic spectrometer VAMOS++ and a new detection arm close to the target. The average fragment N/Z ratio and prompt neutron M_{n} multiplicity are derived and compared with current knowledge from actinide fission. A striking consistency emerges, revealing the unexpected dominant role of the proton subsystem with atomic number between the Z=28 and 50 magic numbers. The origin of nuclear charge polarization in fission and fragment deformation at scission are discussed.
RESUMEN
In the last two decades, through technological, experimental and theoretical advances, the situation in experimental fission studies has changed dramatically. With the use of advanced production and detection techniques both much more detailed and precise information can now be obtained for the traditional regions of fission research and, crucially, new regions of nuclei have become routinely accessible for fission studies. This work first of all reviews the recent developments in experimental fission techniques, in particular the resurgence of transfer-induced fission reactions with light and heavy ions, the emerging use of inverse-kinematic approaches, both at Coulomb and relativistic energies, and of fission studies with radioactive beams. The emphasis on the fission-fragment mass and charge distributions will be made in this work, though some of the other fission observables, such as prompt neutron and γ-ray emission will also be reviewed. A particular attention will be given to the low-energy fission in the so far scarcely explored nuclei in the very neutron-deficient lead region. They recently became the focus for several complementary experimental studies, such as ß-delayed fission with radioactive beams at ISOLDE(CERN), Coulex-induced fission of relativistic secondary beams at FRS(GSI), and several prompt fusion-fission studies. The synergy of these approaches allows a unique insight in the new region of asymmetric fission around [Formula: see text]Hg, recently discovered at ISOLDE. Recent extensive theoretical efforts in this region will also be outlined. The unprecedented high-quality data for fission fragments, completely identified in Z and A, by means of reactions in inverse kinematics at FRS(GSI) and VAMOS(GANIL) will be also reviewed. These experiments explored an extended range of mercury-to-californium elements, spanning from the neutron-deficient to neutron-rich nuclides, and covering both asymmetric, symmetric and transitional fission regions. Some aspects of heavy-ion induced fusion-fission and quasifission reactions will be also discussed, which reveal their dynamical features, such as the fission time scale. The crucial role of the multi-chance fission, probed by means of multinucleon-transfer induced fission reactions, will be highlighted. The review will conclude with the discussion of the new experimental fission facilities which are presently being brought into operation, along with promising 'next-generation' fission approaches, which might become available within the next decade.
RESUMEN
This Letter reports on a systematic study of ß-decay half-lives of neutron-rich nuclei around doubly magic (208)Pb. The lifetimes of the 126-neutron shell isotone (204)Pt and the neighboring (200-202)Ir, (203)Pt, (204)Au are presented together with other 19 half-lives measured during the "stopped beam" campaign of the rare isotope investigations at GSI collaboration. The results constrain the main nuclear theories used in calculations of r-process nucleosynthesis. Predictions based on a statistical macroscopic description of the first-forbidden ß strength reveal significant deviations for most of the nuclei with N<126. In contrast, theories including a fully microscopic treatment of allowed and first-forbidden transitions reproduce more satisfactorily the trend in the measured half-lives for the nuclei in this region, where the r-process pathway passes through during ß decay back to stability.
RESUMEN
We have previously shown that benzamidine-type compounds can inhibit the activity of arginine-specific cysteine proteinases (gingipains HRgpA and RgpB); well-known virulence factors of Porphyromonas gingivalis. They also hinder in vitro growth of this important periodontopathogenic bacterium. Apparently growth arrest is not associated with their ability to inhibit these proteases, because pentamidine, which is a 20-fold less efficient inhibitor of gingipain than 2,6-bis-(4-amidinobenzyl)-cyclohexanone (ACH), blocked P. gingivalis growth far more effectively. To identify targets for benzamidine-derived compounds other than Arg-gingipains, and to explain their bacteriostatic effects, P. gingivalis ATCC 33277 and P. gingivalis M5-1-2 (clinical isolate) cell extracts were subjected to affinity chromatography using a benzamidine-Sepharose column to identify proteins interacting with benzamidine. In addition to HRgpA and RgpB the analysis revealed heat-shock protein GroEL as another ligand for benzamidine. To better understand the effect of benzamidine-derived compounds on P. gingivalis, bacteria were exposed to benzamidine, pentamidine, ACH and heat, and the expression of gingipains and GroEL was determined. Exposure to heat and benzamidine-derived compounds caused significant increases in GroEL, at both the mRNA and protein levels. Interestingly, despite the fact that gingipains were shown to be the main virulence factors in a fertilized egg model of infection, mortality rates were strongly reduced, not only by ACH, but also by pentamidine, a relatively weak gingipain inhibitor. This effect may depend not only on gingipain inhibition but also on interaction of benzamidine derivatives with GroEL. Therefore these compounds may find use in supportive periodontitis treatment.
Asunto(s)
Benzamidinas/farmacología , Ciclohexanonas/farmacología , Pentamidina/farmacología , Porphyromonas gingivalis/efectos de los fármacos , Porphyromonas gingivalis/patogenicidad , Adhesinas Bacterianas/biosíntesis , Adhesinas Bacterianas/genética , Animales , Chaperonina 60/antagonistas & inhibidores , Chaperonina 60/biosíntesis , Chaperonina 60/genética , Embrión de Pollo , Cromatografía de Afinidad , Cisteína Endopeptidasas/biosíntesis , Cisteína Endopeptidasas/genética , Cisteína-Endopeptidasas Gingipaínas , Calor , Virulencia/efectos de los fármacos , Factores de Virulencia/antagonistas & inhibidores , Factores de Virulencia/biosíntesisRESUMEN
Based on a new model of productivity in age diverse tams, findings from a six-year research program are reported in which data from more than 745 natural teams with 8,848 employees in three different fields (car production, administrative work, financial services) were collected. Moreover, central assumptions of this model were tested with a representative survey of the German workforce (N = 2,000). Results support both significant advantages and disadvantages for age-mixed teams. Based on the findings, the following preconditions for the effectiveness of age diverse teams are identified: high task complexity, low salience and high appreciation of age diversity, a positive team climate, low age-discrimination, ergonomic design of work places, and the use of age differentiated leadership. Based on these insights, we developed a new training for supervisors, which addresses the aforementioned aspects and seeks to improve team performance and health of team members. It was found that the training reduces age stereotypes, team conflicts and enhances innovation. Thus, we can conclude that effective interventions for a successful integration of elderly employees in work groups are available and that combinations of measures that address ergonomic design issues, team composition and leadership are to be strongly recommended for practice.
Asunto(s)
Distribución por Edad , Procesos de Grupo , Conducta Cooperativa , Ergonomía , Humanos , Relaciones Interpersonales , Liderazgo , Evaluación de Programas y Proyectos de SaludRESUMEN
To further substantiate gestational age-related changes in oxalate excretion, we studied urinary oxalate excretion in 66 preterm infants born at 23.4-34.7 weeks of gestation. Spot urine of 66 preterm infants was analysed by ion chromatography as soon as they were completely orally fed with enriched breast milk and/or special preterm milk formula (days 7 to 57 of postnatal life). Infants with evidence of renal, gastrointestinal, muscular or metabolic disease were not included. Newborns on parenteral nutrition were excluded. Oxalate/creatinine ratios (Ox/Cr) decreased with gestational age (three age groups: group 1, 23 0/7-28 0/7; group 2, 28 1/7-32 0/7; and group 3, 32 1/7-35 0/7 weeks of gestation). The mean Ox/Cr was highest in group 1 (398.2 mmol/mol +/- 116.8; n = 21). Differences between groups 1 + 3 were statistically significant; p = 0.001; those between groups 1 + 2 and between groups 2 + 3 were not. Ox/Cr correlated inversely with gestational and maturational age (r = -0.41, p = 0.001; r = -0.33, p = 0.007) and positively with postnatal age (r = 0.32, p = 0.008). It correlated inversely with birth weight as well as actual weight at sample collection (r = -0.46 and -0.44, p < 0.001). Ox/Cr was significantly linked to energy and carbohydrate intake (r = 0.3 and 0.4, p = 0.03 and 0.001). These results were independent of sex. In the present study we show that urinary oxalate excretion in preterm infants depends on gestational age.
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Desarrollo Infantil/fisiología , Nutrición Enteral , Trastornos de la Nutrición del Lactante/terapia , Recien Nacido Prematuro/orina , Ácido Oxálico/orina , Estudios de Cohortes , Creatinina/orina , Ingestión de Energía/fisiología , Femenino , Edad Gestacional , Humanos , Fenómenos Fisiológicos Nutricionales del Lactante , Recién Nacido , Recien Nacido Prematuro/crecimiento & desarrollo , Recien Nacido Prematuro/fisiología , Masculino , Factores de TiempoRESUMEN
We report on a novel experimental approach for studying the dissipative spreading of collective motion in a metastable nuclear system, using, for the first time, highly fissile nuclei with spherical shape. This was achieved by fragmentation of 45 radioactive heavy-ion beams at GSI, Darmstadt. The use of inverse kinematics and a dedicated experimental setup allowed for the identification in atomic number of both fission fragments. From the width of their nuclear-charge distributions, a transient time of (3.3+/-0.7)x10(-21) s is deduced for initially spherical nuclei.
RESUMEN
Molecular biological methods such as polymerase chain reaction (PCR) enable microbiologists to detect bacteria even if antibiotic treatment has already been started. Based on this case report of a 39-year-old man with multiple intracerebral lesions, we describe a PCR method called 16S rDNA-PCR which can be used to identify panbacterial DNA by focussing on the universal gene sequences for the bacterial 16S part of the ribosome.
Asunto(s)
Absceso Encefálico/diagnóstico , ADN Bacteriano/genética , Infecciones por Fusobacterium/diagnóstico , Fusobacterium nucleatum/genética , Reacción en Cadena de la Polimerasa/métodos , ARN Ribosómico 16S/genética , Adulto , Encéfalo/patología , Absceso Encefálico/tratamiento farmacológico , Absceso Encefálico/microbiología , ADN Ribosómico , Diagnóstico Diferencial , Infecciones por Fusobacterium/microbiología , Humanos , Imagen por Resonancia Magnética , Masculino , Sensibilidad y Especificidad , Análisis de Secuencia de ADNRESUMEN
BACKGROUND/AIMS: The purpose of the study was to investigate the intracellular survival of Porphyromonas gingivalis as a possible mechanism for maintaining periodontitis. METHODS: P. gingivalis strains, the strain ATCC 33277 and seven clinical isolates, were co-cultured with KB cells. The number of intracellular bacteria was determined up to 3 days after infection. In addition, the numbers of KB cells per well, the concentrations of the cytokines interleukin-1beta (IL-1beta), IL-6, IL-8 and tumour necrosis factor-alpha (TNF-alpha) and the arginine-specific amidolytic activity were measured. The 16S rRNA of P. gingivalis and the mRNA expression of IL-1beta, IL-6, IL-8, TNF-alpha and rgpA were also determined. RESULTS: All the P. gingivalis strains studied were able to survive within KB cells. In contrast to the reduced values of colony-forming units at day 3, equal and higher levels of 16S rRNA were seen in comparison to day 0. Arginine-specific amidolytic activity declined in all samples during infection. Expression of mRNA for rgpA was not found after infection of KB cells by P. gingivalis strains. IL-8 was detectable in all samples 2 days after infection with P. gingivalis strains. Principal components analysis underlined a correlation between the arginine-specific amidolytic activity 1 h after infection and both the released IL-8 and the mRNA expression of IL-8. Associations were found between the cultivable numbers of intracellular P. gingivalis and the mRNAs of IL-1, IL-6 and TNF-alpha at the day of infection. CONCLUSION: The results indicate survival of P. gingivalis within epithelial cells, possibly in a non-cultivable stage. Invasion into cells modulates the virulence properties of P. gingivalis as well as the inflammatory response of the cells.
Asunto(s)
Células KB/microbiología , Porphyromonas gingivalis/fisiología , Adhesinas Bacterianas/análisis , Recuento de Células , Técnicas de Cocultivo , Recuento de Colonia Microbiana , Cisteína Endopeptidasas/análisis , Cisteína-Endopeptidasas Gingipaínas , Humanos , Interleucina-1/análisis , Interleucina-1/genética , Interleucina-6/análisis , Interleucina-6/genética , Interleucina-8/análisis , Interleucina-8/genética , Espacio Intracelular/microbiología , Células KB/inmunología , Porphyromonas gingivalis/enzimología , Porphyromonas gingivalis/inmunología , ARN Mensajero/análisis , ARN Ribosómico 16S/análisis , Factores de Tiempo , Factor de Necrosis Tumoral alfa/análisis , Factor de Necrosis Tumoral alfa/genética , VirulenciaRESUMEN
Spallation residues and fission fragments from 1A GeV 238U projectiles irradiating a liquid hydrogen target were investigated by using the fragment separator at GSI for magnetic selection of reaction products including ray-tracing, energy-loss and time-of-flight techniques. The longitudinal-momentum spectra of identified fragments were analyzed, and evaporation residues and fission fragments could be separated. For 1385 nuclides, production cross sections down to values of 10 microb with a mean accuracy of 15%, velocities in the uranium rest frame and kinetic energies were determined. In the reaction all elements from uranium to nitrogen were found, each with a large number of isotopes.
RESUMEN
A new experimental approach is introduced to investigate the relaxation of the nuclear deformation degrees of freedom. Highly excited fissioning systems with compact shapes and low angular momenta are produced in peripheral relativistic heavy-ion collisions. Both fission fragments are identified in atomic number. Fission cross sections and fission-fragment element distributions are determined as a function of the fissioning element. From the comparison of these new observables with a nuclear-reaction code a value for the transient time is deduced.
RESUMEN
Mechanisms for somatic hypermutation (SHM) have proven elusive. An actively transcribed substrate was analyzed to elucidate the role of stem-loop structures (SLSs) in SHM. Analysis with a new computer algorithm indicates that the location and mutability of a base are regulated by: (a) the extent to which it is unpaired, (b) the degree to which it is exposed by stabilization of SLSs containing and flanking it, and (c) the level of transcription that drives supercoiling, which creates and stabilizes SLSs containing unpaired bases vulnerable to mutation. New mechanisms are described by which transcription can differentially stabilize SLSs harboring targeted bases and establish specific base exposure patterns. Assuming that transcription levels correlate with the magnitude of superhelicity induced and the lengths of ssDNA forming SLSs, this analysis accounts for the location of all mutable bases during SHM.
Asunto(s)
ADN/química , ADN/genética , Genes p53/genética , Mutación , Conformación de Ácido Nucleico , Transcripción Genética , Composición de Base , Secuencia de Bases , Escherichia coli/genética , Proteínas de Escherichia coli/genética , Regulación Bacteriana de la Expresión Génica , Modelos Genéticos , Datos de Secuencia MolecularRESUMEN
Precise momentum distributions of identified projectile fragments, formed in the reactions 238U+Pb and 238U+Ti at 1A GeV, are measured with a high-resolution magnetic spectrometer. With increasing mass loss, the velocities first decrease as expected from previously established systematics, then level off, and finally increase again. Light fragments are on the average even faster than the projectiles. This finding is interpreted as the response of the spectators to the participant blast. The reacceleration of projectile spectators is sensitive to the nuclear mean field and provides a new tool for investigating the equation of state of nuclear matter.
RESUMEN
A total of 305 Ixodes ricinus ticks collected from three areas of Thuringia in central Germany were investigated for infection with Borrelia burgdorferi sensu lato species and Anaplasma phagocytophila. Overall, 11.1% were infected with Borrelia burgdorferi and 2.3% with Anaplasma phagocytophila. Adult ticks showed a significantly higher rate of infection with both borreliae and Anaplasma phagocytophila. Borrelia garinii (55.9%) was detected most frequently, followed by Borrelia burgdorferi sensu stricto (32.4%), Borrelia afzelii (17.6%), and Borrelia valaisiana (5.9%). Four ticks had dual infection with Borrelia garinii and Borrelia burgdorferi sensu stricto. Two of the Borrelia-positive ticks were coinfected with Anaplasma phagocytophila.
Asunto(s)
Anaplasma/aislamiento & purificación , Grupo Borrelia Burgdorferi/aislamiento & purificación , Ixodes/microbiología , Enfermedad de Lyme/epidemiología , Enfermedad de Lyme/microbiología , Infestaciones por Garrapatas/epidemiología , Adulto , Distribución por Edad , Animales , Comorbilidad , ADN Bacteriano/análisis , Femenino , Estudios de Seguimiento , Alemania/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Prevalencia , Medición de Riesgo , Índice de Severidad de la Enfermedad , Distribución por Sexo , Infestaciones por Garrapatas/diagnósticoRESUMEN
In this study we have investigated whether streptolysin O contributes to the virulence of group A streptococci. For this purpose we generated M-negative and SLO-negative mutants by insertion mutagenesis into the chromosome of an M type 1 strain. The inactivation of M1 protein expression was achieved by the construction of the integrative plasmid pSFABS, which contains the internal fragment abs of the emm1 gene. Integration of pSFABS by homologous recombination into the chromosome of strain 38 541 resulted in the generation of mutant EMM1. Inactivation of slo with plasmid pFWSLOD resulted in two different mutant forms. The homologous recombination with plasmid pFWSLOD carrying the two slo fragments slo1 (899 base pairs in the 5' region) and slo2 (709 base pairs in the downstream part) resulted in mutants SLO3, SLO4 and SLO17. In SLO17 a double crossover event took place with insertion of the spectinomycin resistance gene aad9 between the slo fragments 1 and 2. In mutants SLO3 and SLO4 the homologous recombination with the same plasmid led to the integration of the whole plasmid construct into the chromosome of strain 38 541. Both forms of mutation failed to express SLO. In mutant SLO4 additionally M1 protein expression was significantly decreased. The mutants EMM1 (M-, SLO+) and SLO4 (M decreased, SLO-) showed a reduced binding to collagen-coated surfaces. In contrast the mutants SLO3 and SLO17 (both M+, SLO-) and the wild-type strain 38 541 (M+, SLO+) showed an affinity to collagen similar to purified M1 protein. All mutants were less virulent for chicken embryos compared to the wild-type strain after infection by intravenous injection as well as by application onto the chorioallantoic membrane. The results show that besides M protein SLO can also influence virulence of group A streptococci. Moreover, it became obvious that streptococci need more than one tool to fully develop their infectious potential.
Asunto(s)
Antígenos Bacterianos , Proteínas de la Membrana Bacteriana Externa/toxicidad , Proteínas Portadoras/toxicidad , Streptococcus pyogenes/patogenicidad , Estreptolisinas/toxicidad , Animales , Adhesión Bacteriana , Proteínas Bacterianas , Embrión de Pollo , Colágeno/fisiología , Mutación , VirulenciaRESUMEN
The mitogenic factor (MF) of group A streptococci has been reported to be a superantigen stimulating human T cells carrying Vbeta2, 4 and 8 and has been designated streptococcal pyrogenic exotoxin F (SPEF). MF was also shown to possess DNase activity. Here we have purified MF from culture supernatants of different Streptococcus pyogenes strains. Surprisingly, the MF preparations from different strains showed different Vbeta specificities depending on the expression of SPEC or SMEZ3 by the producing strain. Their mitogenic activity decreased upon further purification. In addition, the mitogenic activity could be only neutralized by antibodies against the basic streptococcal superantigens SPEC or SPEX (SMEZ3) but not by antibodies against MF itself although the latter were able to neutralize completely the DNase activity of MF. We found that streptodornase type B (SDB) was expressed in two molecular forms (SDBI and SDBII), differing only by one additional N-terminal arginine at SDBI. MF was found identical to the enzyme SDBII but is devoid of superantigenic properties and should no longer be called a superantigen or a pyrogenic exotoxin.
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Toxinas Bacterianas/inmunología , Desoxirribonucleasa I/inmunología , Exotoxinas/inmunología , Interleucina-2/inmunología , Streptococcus pyogenes/inmunología , Superantígenos/inmunología , Anticuerpos Antibacterianos/inmunología , Desoxirribonucleasa I/antagonistas & inhibidores , Desoxirribonucleasa I/aislamiento & purificación , Humanos , Interleucina-2/aislamiento & purificación , Activación de Linfocitos , Pruebas de Neutralización , Streptococcus pyogenes/genética , Superantígenos/aislamiento & purificación , Linfocitos T/inmunologíaRESUMEN
Spallation residues produced in 1 GeV per nucleon 208Pb on proton reactions have been studied using the Fragment Separator facility at GSI. Isotopic production cross sections of elements from 61Pm to 82Pb have been measured down to 0.1 mb with a high accuracy. The recoil kinetic energies of the produced fragments were also determined. The obtained cross sections agree with most of the few existing gamma-spectroscopic data. The data are compared with different intranuclear-cascade and evaporation-fission models. Drastic deviations were found for a standard code used in technical applications.
RESUMEN
Chlamydia pneumoniae infection has been associated with asthma and atherosclerosis. Smooth muscle cells represent host cells for chlamydiae during chronic infection. In this study we demonstrated that C. pneumoniae infection of human smooth muscle cells in vitro increased production of interleukin 6 (IL-6) and basic fibroblast growth factor (bFGF) as shown by reverse transcription-PCR, immunoblotting, and enzyme-linked immunosorbent assay. In contrast, levels of platelet-derived growth factor A-chain mRNA were not affected after infection. The stimulation of bFGF and IL-6 production was most effective when viable chlamydiae were used as inoculum. Furthermore, inhibition of bacterial protein synthesis with chloramphenicol prevented up-regulation of IL-6 and bFGF in infected cells. Addition of IL-6 antibody to infected cultures diminished bFGF expression, indicating involvement of produced IL-6. These findings suggest that chlamydial infection of smooth muscle cells elicits a cytokine response that may contribute to structural remodeling of the airway wall in chronic asthma and to fibrous plaque formation in atherosclerosis.
Asunto(s)
Chlamydophila pneumoniae/inmunología , Factor 2 de Crecimiento de Fibroblastos/biosíntesis , Interleucina-6/biosíntesis , Músculo Liso/microbiología , Arteriosclerosis/etiología , Asma/etiología , Bronquios/citología , Chlamydia trachomatis/inmunología , Inmunoensayo , Factor de Crecimiento Derivado de Plaquetas/biosíntesis , Especificidad de la EspecieRESUMEN
In addition to beta-haemolytic streptococci belonging to Lancefield group A (Streptococcus pyogenes, GAS), human isolates of group C (GCS) and group G (GGS) streptococci (S. dysgalactiae subsp. equisimilis) have been implicated as causative agents in outbreaks of purulent pharyngitis, of wound infections and recently also of streptococcal toxic shock-like syndrome. Very little is known about the organisation of the genomic region in which the emm gene of GCS and GGS is located. We have investigated the genome sequences flanking the emm gene in GCS by sequencing neighbouring fragments obtained by inverse PCR. Our sequence data for GCS strains 25287 and H46A revealed two types of arrangement in the emm region, which differ significantly from the known types of mga regulon in GAS. We named this segment of the genome mgrC (for multigene regulon-like segment in group C streptococci). In strains belonging to the first mgrC type (prototype strain 25287) the emm gene is flanked up-stream by mgc, a gene that is 61% identical to the mga gene of GAS. A phylogenetic analysis of the deduced protein sequences showed that Mgc is related to Mga proteins of various types of GAS but forms a distinct cluster. Downstream of emm, the mgrC sequence region is bordered by rel. This gene encodes a protein that functions in the synthesis and degradation of guanosine 3',5' bipyrophosphate (ppGpp) during the stringent regulatory response to amino acid deprivation. In the second mgrC type (prototype strain H46A), the genes mgc and emm are arranged as in type 1. But an additional ORF (orf) is inserted in opposite orientation between emm and rel. This orf shows sequence homology to cpdB, which is present in various microorganisms and encodes 2',3' cyclo-nucleotide 2'-phosphodiesterase. PCR analysis showed that these two mgrC arrangements also exist in GGS. Our sequence and PCR data further showed that both types of mgrC region in GCS and GGS are linked via rel to the streptokinase region characterised recently in strain H46A. A gene encoding C5a peptidase, which is present at the 3' end of the mga regulon in GAS, was not found in the mgrC region identified in the GCS and GGS strains investigated here.
Asunto(s)
Antígenos Bacterianos , Proteínas de la Membrana Bacteriana Externa , Proteínas Bacterianas/genética , Proteínas Portadoras/genética , Genes Bacterianos , Familia de Multigenes , Streptococcus/genética , Secuencia de Aminoácidos , Secuencia de Bases , Paseo de Cromosoma , Cartilla de ADN/genética , ADN Bacteriano/genética , Genes Reguladores , Genoma Bacteriano , Humanos , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , Homología de Secuencia de Aminoácido , Streptococcus/clasificación , Streptococcus/aislamiento & purificaciónRESUMEN
A 246-bp imperfect palindrome has the potential to form hairpin structures in single-stranded DNA during replication. Genetic evidence suggests that these structures are converted to double-strand breaks by the SbcCD nuclease and that the double-strand breaks are repaired by recombination. We investigated the role of a range of recombination mutations on the viability of cells containing this palindrome. The palindrome was introduced into the Escherichia coli chromosome by phage lambda lysogenization. This was done in both wt and sbcC backgrounds. Repair of the SbcCD-induced double-strand breaks requires a large number of proteins, including the components of both the RecB and RecF pathways. Repair does not involve PriA-dependent replication fork restart, which suggests that the double-strand break occurs after the replication fork has passed the palindrome. In the absence of SbcCD, recombination still occurs, probably using a gap substrate. This process is also PriA independent, suggesting that there is no collapse of the replication fork. In the absence of RecA, the RecQ helicase is required for palindrome viability in a sbcC mutant, suggesting that a helicase-dependent pathway exists to allow replicative bypass of secondary structures.