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1.
Nature ; 611(7935): 265-270, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-36261531

RESUMEN

The visible world is founded on the proton, the only composite building block of matter that is stable in nature. Consequently, understanding the formation of matter relies on explaining the dynamics and the properties of the proton's bound state. A fundamental property of the proton involves the response of the system to an external electromagnetic field. It is characterized by the electromagnetic polarizabilities1 that describe how easily the charge and magnetization distributions inside the system are distorted by the electromagnetic field. Moreover, the generalized polarizabilities2 map out the resulting deformation of the densities in a proton subject to an electromagnetic field. They disclose essential information about the underlying system dynamics and provide a key for decoding the proton structure in terms of the theory of the strong interaction that binds its elementary quark and gluon constituents. Of particular interest is a puzzle in the electric generalized polarizability of the proton that remains unresolved for two decades2. Here we report measurements of the proton's electromagnetic generalized polarizabilities at low four-momentum transfer squared. We show evidence of an anomaly to the behaviour of the proton's electric generalized polarizability that contradicts the predictions of nuclear theory and derive its signature in the spatial distribution of the induced polarization in the proton. The reported measurements suggest the presence of a new, not-yet-understood dynamical mechanism in the proton and present notable challenges to the nuclear theory.

2.
Phys Rev Lett ; 123(18): 182501, 2019 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-31763910

RESUMEN

Backward-angle meson electroproduction above the resonance region, which was previously ignored, is anticipated to offer unique access to the three quark plus sea component of the nucleon wave function. In this Letter, we present the first complete separation of the four electromagnetic structure functions above the resonance region in exclusive ω electroproduction off the proton, ep→e^{'}pω, at central Q^{2} values of 1.60, 2.45 GeV^{2}, at W=2.21 GeV. The results of our pioneering -u≈-u_{min} study demonstrate the existence of a unanticipated backward-angle cross section peak and the feasibility of full L/T/LT/TT separations in this never explored kinematic territory. At Q^{2}=2.45 GeV^{2}, the observed dominance of σ_{T} over σ_{L}, is qualitatively consistent with the collinear QCD description in the near-backward regime, in which the scattering amplitude factorizes into a hard subprocess amplitude and baryon to meson transition distribution amplitudes: universal nonperturbative objects only accessible through backward-angle kinematics.

3.
Ecotoxicology ; 20(6): 1361-7, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-21533775

RESUMEN

Use of pesticides and other agro-chemicals adversely influence amphibians either directly by killing them or by inducing sublethal, chronic effects. Many studies have investigated the effect of mixtures of pesticides or fertilizers. We studied the combined effects of nitrate and malathion ([(dimethoxy phosphino thioyl] butanediotae) on the early growth, expression of abnormalities, and mortality of Wood Frog (Rana sylvatica) tadpoles in a laboratory experiment. Tadpoles were treated with factorial combinations of 0, 8, and 16 mg NO(3)-N l(-1) and 0, 250, 500, and 1,000 µg malathion l(-1) for a period of 14 days. Feeding behaviour, total length, mean tadpole mass, frequencies of abnormalities, and survivorship in each treatment were recorded. Malathion showed a significant negative influence on all parameters and strongly influenced the frequencies of morphological anomalies. In contrast, nitrate alone did not produce any significant effects on behavior, total length, tadpole mass, or the frequency of abnormalities during the experiment. Malathion and nitrate had an interactive effect on tadpole length and mass, but did not affect any other parameters. Our results suggest that exposure to malathion, even at relatively low concentrations can have serious negative consequences for Wood Frog tadpoles. In addition, our results also indicate that there was little synergistic interaction between malathion and nitrate exposure under laboratory conditions.


Asunto(s)
Insecticidas/toxicidad , Malatión/toxicidad , Nitratos/toxicidad , Ranidae/fisiología , Contaminantes Químicos del Agua/toxicidad , Animales , Larva/efectos de los fármacos , Larva/crecimiento & desarrollo , Ranidae/anomalías , Ranidae/crecimiento & desarrollo
4.
Science ; 267(5201): 1166-9, 1995 Feb 24.
Artículo en Inglés | MEDLINE | ID: mdl-7855597

RESUMEN

Exonuclease I (Exo I) from Schizosaccharomyces pombe, a 5'-->3' double-stranded DNA exonuclease, is induced during meiotic prophase I. The exo1 gene is a member of a family of related DNA repair genes, including RAD2/rad13/xpgc and YKL510/rad2, conserved from yeast to humans. An exo1 mutant displays a mutator phenotype and alters activity of the ade6-M387 marker effect. These results suggest that Exo I acts in a pathway that corrects mismatched base pairs.


Asunto(s)
Reparación del ADN , ADN de Hongos/metabolismo , Exodesoxirribonucleasas/metabolismo , Mutación , Schizosaccharomyces/genética , Alelos , Secuencia de Aminoácidos , Composición de Base , Secuencia de Bases , Cruzamientos Genéticos , Replicación del ADN , Exodesoxirribonucleasas/química , Exodesoxirribonucleasas/genética , Meiosis , Datos de Secuencia Molecular , Recombinación Genética , Schizosaccharomyces/enzimología , Schizosaccharomyces/fisiología
5.
Science ; 175(4022): 644-6, 1972 Feb 11.
Artículo en Inglés | MEDLINE | ID: mdl-5009763

RESUMEN

Catostomid fishes appear to have 2n(-->4n?) approximately 100 chromosomes. The Cyprinidae, from which catostomids probably diverged before the Eocene, usually have 2n = 48 or 50 chromosomes. Preliminary cytophotometric measurements indicate an approximate doubling of DNA content of cells among catostomids.


Asunto(s)
Peces , Cariotipificación , Poliploidía , Animales , Evolución Biológica , Cyprinidae , Citogenética , ADN/análisis , Células Epiteliales , Branquias/citología , Mitosis
6.
Science ; 215(4532): 548-53, 1982 Jan 29.
Artículo en Inglés | MEDLINE | ID: mdl-17771276

RESUMEN

Combined analysis of helium (584 angstroms) airglow and the atmospheric occultations of the star delta Scorpii imply a vertical mixing parameter in Saturn's upper atmosphere of K (eddy diffusion coefficient) approximately 8 x 10(7) square centimeters per second, an order of magnitude more vigorous than mixing in Jupiter's upper atmosphere. Atmospheric H(2) band absorption of starlight yields a preliminary temperature of 400 K in the exosphere and a temperature near the homopause of approximately 200 K. The energy source for the mid-latitude H(2) band emission still remains a puzzle. Certain auroral emissions can be fully explained in terms of electron impact on H(2), and auroral morphology suggests a link between the aurora and the Saturn kilometric radiation. Absolute optical depths have been determined for the entire C ring andparts of the A and B rings. A new eccentric ringlet has been detected in the C ring. The extreme ultraviolet reflectance of the rings is fairly uniform at 3.5 to 5 percent. Collisions may control the distribution of H in Titan's H torus, which has a total vertical extent of approximately 14 Saturn radii normal to the orbit plane.

7.
Science ; 233(4759): 74-9, 1986 Jul 04.
Artículo en Inglés | MEDLINE | ID: mdl-17812892

RESUMEN

Data from solar and stellar occultations of Uranus indicate a temperature of about 750 kelvins in the upper levels of the atmosphere (composed mostly of atomic and molecular hydrogen) and define the distributions of methane and acetylene in the lower levels. The ultraviolet spectrum of the sunlit hemisphere is dominated by emissions from atomic and molecular hydrogen, which are kmown as electroglow emissions. The energy source for these emissions is unknown, but the spectrum implies excitation by low-energy electrons (modeled with a 3-electron-volt Maxwellian energy distribution). The major energy sink for the electrons is dissociation of molecular hydrogen, producing hydrogen atoms at a rate of 10(29) per second. Approximately half the atoms have energies higher than the escape energy. The high temperature of the atmosphere, the small size of Uranus, and the number density of hydrogen atoms in the thermosphere imply an extensive thermal hydrogen corona that reduces the orbital lifetime of ring particles and biases the size distribution toward larger particles. This corona is augmented by the nonthermal hydrogen atoms associated with the electroglow. An aurora near the magnetic pole in the dark hemisphere arises from excitation of molecular hydrogen at the level where its vertical column abundance is about 10(20) per square centimeter with input power comparable to that of the sunlit electroglow (approximately 2x10(11) watts). An initial estimate of the acetylene volume mixing ratio, as judged from measurements of the far ultraviolet albedo, is about 2 x 10(-7) at a vertical column abundance of molecular hydrogen of 10(23) per square centimeter (pressure, approximately 0.3 millibar). Carbon emissions from the Uranian atmosphere were also detected.

8.
Science ; 212(4491): 206-11, 1981 Apr 10.
Artículo en Inglés | MEDLINE | ID: mdl-17783831

RESUMEN

The global hydrogen Lyman alpha, helium (584 angstroms), and molecular hydrogen band emissions from Saturn are qualitatively similar to those of Jupiter, but the Saturn observations emphasize that the H(2) band excitation mechanism is closely related to the solar flux. Auroras occur near 80 degrees latitude, suggesting Earth-like magnetotail activity, quite different from the dominant Io plasma torus mechanism at Jupiter. No ion emissions have been detected from the magnetosphere of Saturn, but the rings have a hydrogen atmosphere; atomic hydrogen is also present in a torus between 8 and 25 Saturn radii. Nitrogen emission excited by particles has been detected in the Titan dayglow and bright limb scans. Enhancement of the nitrogen emission is observed in the region of interaction between Titan's atmosphere and the corotating plasma in Saturn's plasmasphere. No particle-excited emission has been detected from the dark atmosphere of Titan. The absorption profile of the atmosphere determined by the solar occultation experiment, combined with constraints from the dayglow observations and temperature information, indicate that N(2) is the dominant species. A double layer structure has been detected above Titan's limb. One of the layers may be related to visible layers in the images of Titan.

9.
Science ; 246(4936): 1459-66, 1989 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-17756000

RESUMEN

Results from the occultation of the sun by Neptune imply a temperature of 750 +/- 150 kelvins in the upper levels of the atmosphere (composed mostly of atomic and molecular hydrogen) and define the distributions of methane, acetylene, and ethane at lower levels. The ultraviolet spectrum of the sunlit atmosphere of Neptune resembles the spectra of the Jupiter, Saturn, and Uranus atmospheres in that it is dominated by the emissions of H Lyman alpha (340 +/- 20 rayleighs) and molecular hydrogen. The extreme ultraviolet emissions in the range from 800 to 1100 angstroms at the four planets visited by Voyager scale approximately as the inverse square of their heliocentric distances. Weak auroral emissions have been tentatively identified on the night side of Neptune. Airglow and occultation observations of Triton's atmosphere show that it is composed mainly of molecular nitrogen, with a trace of methane near the surface. The temperature of Triton's upper atmosphere is 95 +/- 5 kelvins, and the surface pressure is roughly 14 microbars.

10.
Trends Biochem Sci ; 25(8): 368-74, 2000 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-10916155

RESUMEN

Ion channels mediate electrical excitability in neurons and muscle. Three-dimensional structures for model peptide channels and for a potassium (K+) channel have been combined with computer simulations to permit rigorous exploration of structure-function relations of channels. Water molecules and ions within transbilayer pores tend to diffuse more slowly than in bulk solutions. In the narrow selectivity filter of the bacterial K+ channel (i.e. the region of the channel that discriminates between different species of ions) a column of water molecules and K+ ions moves in a concerted fashion. By combining atomistic simulations (in which all atoms of the channel molecule, water and ions are treated explicitly) with continuum methods (in which the description of the channel system is considerably simplified) it is possible to simulate some of the physiological properties of channels.


Asunto(s)
Alameticina/metabolismo , Gramicidina/metabolismo , Canales Iónicos/fisiología , Canales de Potasio/metabolismo , Estructura Cuaternaria de Proteína , Alameticina/química , Antibacterianos/metabolismo , Membrana Celular/metabolismo , Simulación por Computador , Difusión , Gramicidina/química , Canales Iónicos/química , Proteínas de la Membrana , Modelos Moleculares , Permeabilidad , Canales de Potasio/química , Electricidad Estática , Relación Estructura-Actividad
11.
Evolution ; 61(5): 1176-87, 2007 May.
Artículo en Inglés | MEDLINE | ID: mdl-17492970

RESUMEN

The mass-specific metabolic rate hypothesis of Gillooly and others predicts that DNA mutation and substitution rates are a function of body mass and temperature. We tested this hypothesis with sequence divergences estimated from mtDNA cytochrome b sequences of 54 taxa of cyprinid fish. Branch lengths estimated from a likelihood tree were compared with metabolic rates calculated from body mass and environmental temperatures experienced by those taxa. The problem of unknown age estimates of lineage splitting was avoided by comparing estimated amounts of metabolic activity along phyletic lines leading to pairs of modern taxa from their most recent common ancestor with sequence divergences along those same pairs of phyletic lines. There were significantly more pairs for which the phyletic line with greater genetic change also had the higher metabolic activity, when compared to the prediction of a hypothesis that body mass and temperature are not related to substitution rate.


Asunto(s)
Metabolismo Basal/fisiología , Tamaño Corporal/fisiología , Cyprinidae/genética , ADN Mitocondrial , Evolución Molecular , Temperatura , Animales , Funciones de Verosimilitud , Mutación , Homología de Secuencia de Ácido Nucleico
12.
Artículo en Inglés | MEDLINE | ID: mdl-9752725

RESUMEN

Homologous recombination occurs at high frequency during meiosis and is essential for the proper segregation of chromosomes and the generation of genetic diversity. Meiotic recombination is controlled in numerous ways. In the fission yeast Schizosaccharomyces pombe nutritional starvation induces meiosis and high-level expression of many genes, including numerous recombination (rec) genes, whose products are required for recombination. Accompanying the two meiotic divisions are profound changes in nuclear and chromosomal structure and movement, which may play an important role in meiotic recombination. Although recombination occurs throughout the genome, it occurs at high frequency in some intervals (hotspots) and at low frequency in others (coldspots). The well-characterized hotspot M26 is activated by the Mts1/Mts2 protein; this site and its binding proteins interact with the local chromosomal structure to enhance recombination. A coldspot between the silent mating-type loci is repressed by identified proteins, which may also alter local chromatin. We discuss in detail the rec genes and the possible functions of their products, some but not all of which share homology with other identified proteins. Although some of the rec gene products are required for recombination throughout the genome, others demonstrate regional specificity and are required in certain genomic regions but not in others. Throughout the review contrasts are made with meiotic recombination in the more thoroughly studied budding yeast Saccharomyces cerevisiae.


Asunto(s)
Recombinación Genética , Schizosaccharomyces/citología , Schizosaccharomyces/genética , Núcleo Celular/ultraestructura , Cromosomas Fúngicos/genética , Cromosomas Fúngicos/ultraestructura , Genes Fúngicos , Genes del Tipo Sexual de los Hongos , Meiosis , Mutagénesis , Complejo Sinaptonémico/genética
13.
Biochim Biophys Acta ; 1510(1-2): 1-9, 2001 Feb 09.
Artículo en Inglés | MEDLINE | ID: mdl-11342142

RESUMEN

We have performed simulations of both a single potassium ion and a single sodium ion within the pore of the bacterial potassium channel KcsA. For both ions there is a dehydration energy barrier at the cytoplasmic mouth suggesting that the crystal structure is a closed conformation of the channel. There is a potential energy barrier for a sodium ion in the selectivity filter that is not seen for potassium. Radial distribution functions for both ions with the carbonyl oxygens of the selectivity filter indicate that sodium may interact more tightly with the filter than does potassium. This suggests that the key to the ion selectivity of KcsA is the greater dehydration energy of Na(+) ions, and helps to explain the block of KcsA by internal Na(+) ions.


Asunto(s)
Proteínas Bacterianas/química , Canales de Potasio/química , Potasio/química , Sodio/química , Cationes Monovalentes , Modelos Moleculares , Termodinámica , Agua/química
14.
Biochim Biophys Acta ; 1211(3): 289-300, 1994 Mar 24.
Artículo en Inglés | MEDLINE | ID: mdl-8130262

RESUMEN

The rate of hydrolysis of phosphatidylcholine bilayers by phospholipase A2 may be either enhanced or inhibited by the presence of lysolecithin depending on the experimental conditions examined. To further understand the relationship of lysolecithin to phospholipase A2 activity, the binding of lysolecithin to phospholipase A2 from the venom of Agkistrodon piscivorus piscivorus was examined by fluorescence spectroscopy. The tryptophan emission intensity of the enzyme was enhanced by 70% upon addition of lysolecithin. The binding isotherm for lysolecithin to the phospholipase A2 estimated from the fluorescence change was biphasic, with a clear break in the curve occurring at the critical micelle concentration of the lysolecithin. Several observations suggested that the phospholipase A2 was capable of hydrolyzing the lysolecithin although at a rate far below that of phospholipid hydrolysis. These experiments were repeated using several other species of phospholipase A2, and the results were found to be general among the enzymes except the lys-49 isozyme from A. p. piscivorus which displayed neither the dependence on the critical micelle concentration for binding nor the ability to hydrolyze lysolecithin. These results were used as the basis for a quantitative analysis of enzyme fluorescence changes that occur during the time course of phospholipid hydrolysis and of the mechanism whereby lysolecithin inhibits the hydrolysis of phosphatidylcholine bilayers by phospholipase A2.


Asunto(s)
Lisofosfatidilcolinas/metabolismo , Fosfolipasas A/metabolismo , Agkistrodon , Animales , Venenos de Crotálidos/química , Venenos de Crotálidos/metabolismo , Cinética , Fosfolipasas A/química , Fosfolipasas A2 , Espectrometría de Fluorescencia , Triptófano/química
15.
Biochim Biophys Acta ; 1370(1): 1-7, 1998 Mar 06.
Artículo en Inglés | MEDLINE | ID: mdl-9518528

RESUMEN

A model of the selectivity filter of a voltage-gated K+ (Kv) channel formed by an eight-stranded beta-barrel is compared with physiological properties of the channel. Continuum electrostatic calculations suggest that only two of the eight Asp sidechains at the extracellular mouth of the pore will ionise. A ring of four Tyr sidechains forms the narrowest region of the pore. Molecular dynamic simulations of the potential energy of a K+ ion as translated along the model pore indicate that the two ionised Asp sidechains and the hydroxyl groups of the Tyr sidechains stabilise the partially desolvated ion as it passes through the narrowest region.


Asunto(s)
Simulación por Computador , Modelos Moleculares , Canales de Potasio/metabolismo , Proteínas/metabolismo , Agua/metabolismo , Secuencia de Aminoácidos , Animales , Transporte Iónico , Datos de Secuencia Molecular , Estructura Terciaria de Proteína , Relación Estructura-Actividad , Termodinámica
16.
Circulation ; 100(18): 1917-22, 1999 Nov 02.
Artículo en Inglés | MEDLINE | ID: mdl-10545437

RESUMEN

BACKGROUND: To date, the lack of potent and selective inhibitors has hampered the physiological assessment of modulation of the cardiac slowly activating delayed rectifier current, I(Ks). The present study, using the I(Ks) blocker L-768,673, represents the first in vivo assessment of the cardiac electrophysiological and antiarrhythmic effects of selective I(Ks) blockade. METHODS AND RESULTS: In an anesthetized canine model of recent (8.5+/-0.4 days) anterior myocardial infarction, 0.003 to 0.03 mg/kg L-768,673 IV significantly suppressed electrically induced ventricular tachyarrhythmias and reduced the incidence of lethal arrhythmias precipitated by acute, thrombotically induced posterolateral myocardial ischemia. Antiarrhythmic protection afforded by L-768,673 was accompanied by modest 7% to 10% increases in noninfarct zone ventricular effective refractory period, 3% to 5% increases in infarct zone ventricular effective refractory period, and 4% to 6% increases in QTc interval. In a conscious canine model of healed (3 to 4 weeks) anterior myocardial infarction, ventricular fibrillation was provoked by transient occlusion of the left circumflex coronary artery during submaximal exercise. Pretreatment with 0.03 mg/kg L-768,673 IV elicited a modest 7% increase in QTc, prevented ventricular fibrillation in 5 of 6 animals, and suppressed arrhythmias in 2 additional animals. CONCLUSIONS: The present findings suggest that selective blockade of I(Ks) may be a potentially useful intervention for the prevention of malignant ischemic ventricular arrhythmias.


Asunto(s)
Acetamidas/uso terapéutico , Antiarrítmicos/uso terapéutico , Arritmias Cardíacas/tratamiento farmacológico , Benzodiazepinonas/uso terapéutico , Bloqueo Cardíaco/terapia , Isquemia Miocárdica/tratamiento farmacológico , Disfunción Ventricular/tratamiento farmacológico , Animales , Arritmias Cardíacas/etiología , Modelos Animales de Enfermedad , Perros , Electrocardiografía , Isquemia Miocárdica/complicaciones , Sistema Nervioso Simpático/fisiología , Disfunción Ventricular/etiología
17.
J Mol Biol ; 229(1): 67-78, 1993 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-8380618

RESUMEN

RecBCD enzyme of Escherichia coli unwinds DNA from duplex DNA ends to produce single-stranded DNA, a central intermediate in the major (RecBCD) pathway of homologous recombination. To help elucidate the mechanism of unwinding we studied the complex of RecBCD enzyme bound to duplex DNA ends in the absence of ATP, a cofactor required for unwinding. In this complex the terminal 16 or 17 nucleotides of the 3' terminated strand and the terminal 20 or 21 nucleotides of the 5' terminated strand were protected from DNase I cleavage. u.v.-irradiation of the complex cross-linked the RecB subunit to the 3' terminated strand and the RecC and RecD subunits of the 5' terminated strand. Studies using pyridoxal 5-phosphate, an inhibitor of the RecBCD enzyme, corroborated the cross-linking studies and revealed a conformational change in the enzyme upon binding to DNA. Based on these results and proposals by others, we present a model for the mechanism of DNA unwinding by RecBCD enzyme.


Asunto(s)
ADN Helicasas/metabolismo , Proteínas de Unión al ADN/metabolismo , Proteínas de Escherichia coli , Exodesoxirribonucleasas/metabolismo , Secuencia de Bases , Borohidruros , Exodesoxirribonucleasa V , Exodesoxirribonucleasas/efectos de los fármacos , Exodesoxirribonucleasas/efectos de la radiación , Modelos Genéticos , Datos de Secuencia Molecular , Fosfato de Piridoxal , Rayos Ultravioleta
18.
J Mol Biol ; 211(1): 117-34, 1990 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-2405161

RESUMEN

We tested the hypothesis that RecBCD enzyme of Escherichia coli resolves pre-existing Holliday recombination intermediates by examining the action of the purified enzyme on an open-ended DNA cruciform with limited ability to branch migrate. The enzyme cleaved two strands of the cruciform near its base to produce "recombinant" products, with a marked bias in the direction of cleavage. The two nicks necessary to cleave the cruciform were made separately. Cruciforms whose four termini were blocked by synthetic hairpin-shaped oligonucleotides were not detectably nicked by the enzyme. With one terminus open the enzyme made a nick at the base of the cruciform but not a double-strand cut. With two or more termini open the enzyme made double-strand cuts. We infer that RecBCD enzyme molecules must enter the termini of duplex DNA and approach the cruciform from more than one direction in order to cleave it into recombinant products. Previous results on RecBCD-mediated recombination between phage lambda and lambda dv imply that intracellular RecBCD enzyme can approach pre-existing Holliday junctions from only one direction. We infer that intracellular RecBCD enzyme cannot cleave pre-existing Holliday junctions into recombinants and suggest that the enzyme may cleave Holliday junctions in whose formation it participates.


Asunto(s)
ADN/metabolismo , Exodesoxirribonucleasas/metabolismo , Secuencia de Bases , Electroforesis en Gel de Agar , Electroforesis en Gel Bidimensional , Escherichia coli/enzimología , Escherichia coli/genética , Exodesoxirribonucleasa V , Genotipo , Datos de Secuencia Molecular , Conformación de Ácido Nucleico , Oligonucleótidos/síntesis química , Plásmidos , Secuencias Repetitivas de Ácidos Nucleicos , Mapeo Restrictivo , Especificidad por Sustrato
19.
J Mol Biol ; 189(4): 585-95, 1986 Jun 20.
Artículo en Inglés | MEDLINE | ID: mdl-3783685

RESUMEN

Homologous recombination in Escherichia coli occurs at increased frequency near Chi sites, 5'G-C-T-G-G-T-G-G3'. Cutting of DNA close to the Chi sequence by the E. coli RecBC enzyme is essential to Chi's stimulation of recombination. We have detected Chi-dependent cutting activity in extracts of several genera of terrestrial enteric bacteria (family Enterobacteriaceae) and of two genera of marine enteric bacteria (family Vibrionaceae). More distantly related bacteria had no detectable Chi-dependent cutting activity. These results support the view that recognition of this specific nucleotide sequence as a signal activating recombination has been maintained during the evolution of certain groups of bacteria. We discuss the possibility that other sequences play a similar role in other groups of bacteria.


Asunto(s)
Enterobacteriaceae/genética , Recombinación Genética , Vibrionaceae/genética , Autorradiografía , Secuencia de Bases , ADN Bacteriano/genética , ADN Bacteriano/metabolismo , Electroforesis en Gel de Poliacrilamida , Enterobacteriaceae/metabolismo , Vibrionaceae/metabolismo
20.
J Mol Biol ; 194(4): 747-50, 1987 Apr 20.
Artículo en Inglés | MEDLINE | ID: mdl-2958631

RESUMEN

Chi, 5' G-C-T-G-G-T-G-G 3', stimulates coliphage lambda recombination mediated by the major recombination pathway of Escherichia coli, the RecBC pathway. Purified RecBCD enzyme makes a single strand endonuclease cleavage four to six nucleotides to the 3' side of the chi sequence. Three sequences similar to chi, 5' A-C-T-G-G-T-G-G 3', 5' G-T-T-G-G-T-G-G 3' and 5' G-C-T-A-G-T-G-G 3', have partial recombinational hotspot activity in genetic crosses. We report here that purified RecBCD enzyme preferentially cuts four nucleotides to the right of the two most active chi-like octamers. The degree of cutting correlated with the genetic activities of these sequences; this result indicates that these cleavages are essential to the genetic activity of chi and chi-like sequences.


Asunto(s)
Proteínas de Escherichia coli , Escherichia coli/enzimología , Exodesoxirribonucleasas/metabolismo , Recombinación Genética , Bacteriófago lambda/genética , Secuencia de Bases , ADN Viral , Escherichia coli/genética , Exodesoxirribonucleasa V
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