Investigation of EscA as a chaperone for the Edwardsiella tarda type III secretion system putative translocon component EseC

Edwardsiella tarda is an important Gram-negative enteric pathogen affecting both animals and humans. It possesses a type III secretion system (T3SS) essential for pathogenesis. EseB, EseC and EseD have been shown to form a translocon complex after secretion, while EscC functions as a T3SS chaperone for EseB and EseD. In this paper we identify EscA, a protein required for accumulation and proper secretion of another translocon component, EseC. The escA gene is located upstream of eseC and the EscA protein has the characteristics of T3SS chaperones. Cell fractionation experiments indicated that EscA is located in the cytoplasm and on the cytoplasmic membrane. Mutation with in-frame deletion of escA greatly decreased the secretion of EseC, while complementation of escA restored the wild-type secretion phenotype. The stabilization and accumulation of EseC in the cytoplasm were also affected in the absence of EscA. Mutation of escA did not affect the transcription of eseC but reduced the accumulation level of EseC as measured by using an EseC-LacZ fusion protein in Ed. tarda. Co-purification and co-immunoprecipitation studies demonstrated a specific interaction between EscA and EseC. Further analysis showed that residues 31-137 of EseC are required for EseC-EscA interaction, Mutation of EseC residues 31-137 reduced the secretion and accumulation of EseC in Ed. tarda. Finally, infection experiments showed that mutations of EscA and residues 31-137 of EseC increased the LD50 by approximately 10-fold in blue gourami fish. These results indicated that EscA functions as a specific chaperone for EseC and contributes to the virulence of Ed. tarda.

EscC is a chaperone for the Edwardsiella tarda type III secretion system putative translocon components EseB and EseD

Edwardsiella tarda is a Gram-negative enteric pathogen that causes disease in both humans and animals. Recently, a type III secretion system (T3SS) has been found to contribute to Ed. tarda pathogenesis. EseB, EseC and EseD were shown to be secreted by the T3SS and to be the major components of the extracellular proteins (ECPs). Based on sequence similarity, they have been proposed to function as the 'translocon' of the T3SS needle structure. In this study, it was shown that EseB, EseC and EseD formed a protein complex after secretion, which is consistent with their possible roles as translocon components. The secretion of EseB and EseD was dependent on EscC (previously named Orf2). EscC has the characteristics of a chaperone; it is a small protein (13 kDa), located next to the translocators in the T3SS gene cluster, and has a coiled-coil structure at the N-terminal region as predicted by COILS. An in-frame deletion of escC abolished the secretion of EseB and EseD, and complementation of Delta escC restored the export of EseB and EseD into the culture supernatant. Further studies showed that EscC is not a secreted protein and is located on the membrane and in the cytoplasm. Mutation of escC did not affect the transcription of eseB but reduced the amount of EseB as measured by using an EseB-LacZ fusion protein in Ed. tarda. Co-purification studies demonstrated that EscC formed complexes with EseB and EseD. The results suggest that EscC functions as a T3SS chaperone for the putative translocon components EseB and EseD in Ed. tarda.

Phase control of gain and dispersion in an open lambda-type inversionless lasing system

The control role of the relative phase between the probe and driving fields on the gain and dispersion in an open Lambda-type inversionless lasing system with spontaneously generated coherence (SGC) is investigated. It is shown that the inversionless gain and dispersion are quite sensitive to variation in the relative phase; by adjusting the value of the relative phase, electromagnetically induced transparency (EIT), a high refractive index with zero absorption and a larger inversionless gain can be realized. It is also shown that, in the contributions to the inversionless gain ( absorption) and dispersion, the contribution from SGC is always much larger than that from the dynamically induced coherence for any value of the relative phase. Our analysis shows that variation in the SGC effect will cause the spectrum regions and values of the inversionless gain and dispersion to vary evidently. We also found that, under the same conditions, the values of the inversionless gain and dispersion in the open system are evidently larger than those in the corresponding closed system; EIT occurs in the open system but cannot occur in the closed system.

Propagation of a few-cycle laser pulse in a V-type three-level system

Propagation of a few-cycle laser pulse in a V-type three-level system (fine structure levels of rubidium) is investigated numerically. The full three-level Maxwell-Bloch equations without the rotating wave approximation and the standing slowly varying envelope approximation are solved by using a finite-difference time-domain method. It is shown that, when the usual unequal oscillator strengths are considered, self-induced transparency cannot be recovered and higher spectral components can be produced even for small-area pulses. (c) 2005 Pleiades Publishing, Inc.

Six-wave mixing phase-dispersion by optical heterodyne detection in dressed reverse N-type four-level system

We have investigated the dressed effects of non-degenerate four-wave mixing (NDFWM) and demonstrated a phase-sensitive method of studying the fifth-order nonlinear susceptibility due to atomic coherence in RN-type four-level system. In the presence of a strong coupling field, NDFWM spectrum exhibits Autler-Townes splitting, accompanied by either suppression or enhancement of the NDFWM signal, which is directly related to the competition between the absorption and dispersion contributions. The heterodyne-detected nonlinear absorption and dispersion of six-wave mixing signal in the RN-type system show that the hybrid radiation-matter detuning damping oscillation is in the THz range and can be controlled and modified through the colour-locked correlation of twin noisy fields.

Analysis of the effect of copper on the virulence of a pathogenic Edwardsiella tarda strain

Aim: To investigate the effect of copper on the virulence of Edwardsiella tarda. Methods and Results: The pathogenic Edw. tarda strain TX5 was cultured under copper-stressed conditions and examined for any potential alteration in capacities that are associated with pathogenicity. The results showed that compared to untreated TX5, Cu-treated TX5 exhibits reduced planktonic and biofilm growth, an impaired ability to adhere to host mucus, modulation of host immune response, and dissemination in host blood and liver. Consistent with these observations, the overall bacterial virulence of Cu-treated TX5 is significantly attenuated. SDS-PAGE analyses of whole cell protein production showed that Cu-treated TX5 differs from the untreated TX5 in its production of at least one protein. Quantitative real time reverse transcriptase PCR analyses showed that copper treatment decreased the expression of virulence-associated genes encoding components of the type III and type VI secretion systems, the Eth haemolysin system, and the LuxS/AI-2 quorum-sensing system. Conclusions: Prolonged exposure to copper has multiple effects on TX5 and results in significant attenuation of bacterial virulence. Significance and Impact of the Study: The results of this study demonstrate that copper treatment has a broad and profound effect on the virulence-associated capacities of TX5, which is exerted at least in part at the transcription level. These findings provide new insights to the antimicrobial mechanism of copper.

Regulation of autoinducer 2 production and luxS expression in a pathogenic Edwardsiella tarda strain

Edwardsiella tarda is a bacterial pathogen that can infect both humans and animals. TX1, an Ed. tarda strain isolated from diseased fish, was found to produce autoinducer 2 (Al-2)-like activity that was growth phase dependent and modulated by growth conditions. The gene coding for the Al-2 synthase was cloned from TX1 and designated luxS(Et). LuxS(Et) was able to complement the Al-2 mutant phenotype of Escherichia coli strain DH5 alpha. Expression Of luxS(Et) correlated with Al-2 activity and was increased by glucose and decreased by elevated temperature. The effect of glucose was shown to be mediated through the cAMP-CRP complex, which repressed luxS(Et) expression. Overexpression of luxS(Et) enhanced Al-2 activity in TX1, whereas disruption of luxS(Et) expression by antisense RNA interference (i) reduced the level of Al-2 activity, (ii) impaired bacterial growth under various conditions, (iii) weakened the expression of genes associated with the type III secretion system and biofilm formation, and (iv) attenuated bacterial virulence. Addition of exogenous Al-2 was able to complement the deficiencies in the expression of TTSS genes and biofilm production but failed to rescue the growth defects. Our results (i) demonstrated that the Al-2 activity in TX1 is controlled at least in part at the level of luxS(Et) expression, which in turn is regulated by growth conditions, and that the temporal expression of luxS(Et) is essential for optimal bacterial infection and survival; and (ii) suggested the existence in Ed. tarda of a LuxS/Al-2-mediated signal transduction pathway that regulates the production of virulence-associated elements.

Regulation of autoinducer 2 production and luxS expression in a pathogenic Edwardsiella tarda strain

Edwardsiella tarda is a bacterial pathogen that can infect both humans and animals. TX1, an Ed. tarda strain isolated from diseased fish, was found to produce autoinducer 2 (Al-2)-like activity that was growth phase dependent and modulated by growth conditions. The gene coding for the Al-2 synthase was cloned from TX1 and designated luxS(Et). LuxS(Et) was able to complement the Al-2 mutant phenotype of Escherichia coli strain DH5 alpha. Expression Of luxS(Et) correlated with Al-2 activity and was increased by glucose and decreased by elevated temperature. The effect of glucose was shown to be mediated through the cAMP-CRP complex, which repressed luxS(Et) expression. Overexpression of luxS(Et) enhanced Al-2 activity in TX1, whereas disruption of luxS(Et) expression by antisense RNA interference (i) reduced the level of Al-2 activity, (ii) impaired bacterial growth under various conditions, (iii) weakened the expression of genes associated with the type III secretion system and biofilm formation, and (iv) attenuated bacterial virulence. Addition of exogenous Al-2 was able to complement the deficiencies in the expression of TTSS genes and biofilm production but failed to rescue the growth defects. Our results (i) demonstrated that the Al-2 activity in TX1 is controlled at least in part at the level of luxS(Et) expression, which in turn is regulated by growth conditions, and that the temporal expression of luxS(Et) is essential for optimal bacterial infection and survival; and (ii) suggested the existence in Ed. tarda of a LuxS/Al-2-mediated signal transduction pathway that regulates the production of virulence-associated elements.

Construction and characterization of a live, attenuated esrB mutant of Edwardsiella tarda and its potential as a vaccine against the haemorrhagic septicaemia in turbot, Scophthamus maximus (L.)

The esrB gene of Edwardsiella tarda, which encodes a regulator protein of the type III secretion system, was mutated by the unmarked deletion method and reintroduced by allelic exchange into the chromosome of E. tarda LSE40 by means of the suicide vector pRE 112. The LSE40 esrB mutant was highly attenuated when inoculated intraperitoneally into turbot Scophthamus maximus L., showing a 50% lethal dose of 10(8.1) cfu/fish. The esrB mutants were not recoverable from the internal organs at 14 days post-inoculation. Vaccination with a single dose of 10(5)-10(7) cfu/fish of the esrB mutant elicited significant protection against the wildtype strain of E. tarda LSE40 (relative percentage survival > 50%). The protection correlated well with the antibody titres in the serum of vaccinated fish. (c) 2006 Elsevier Ltd. All rights reserved.

Phase control of gain and dispersion in an open V-type system

The control role of the relative phase between the probe and driving fields on gain, dispersion and populations in an open V-type three-level system with spontaneously generated coherence is studied. The result shows that by adjusting the value of the relative phase, the transformation between lasing with inversion and lasing without inversion (LWI) can be realized and high dispersion (refractive index) without absorption can be obtained. The shape and value range of the dispersion curve are similar to those of the gain curve, and this similarity is closely related to the relative phase. The effects of the atomic exit and injection rates and the incoherent pump rate on the control role of the relative phase are also analysed. It is found easier to get LWI by adjusting the value of the relative phase using the open system rather than the closed system, and using an incoherent pump rather than without using the incoherent pump. Moreover the open system can give a larger LWI gain than the closed system.

Dressed multi-wave mixing in a V-type four-level atomic system

The dressed four- and six-wave mixings in a V-type four-level system are considered. Under two different dressed conditions, two- and three-photon resonant Autler-Townes splittings, accompanied by enhancement and suppression of wave mixing signal, are obtained analytically. Meanwhile, an electromagnetic induced transparency of multi-wave mixing is presented, which shows multiple peaks and asymmetric effects caused by one-photon, two-photon and three-photon resonances, separately. The slow light propagation multiple region of multi-wave mixing signal is also obtained.

A multi-standard active-RC filter with accurate tuning system

A low-power, highly linear, multi-standard, active-RC filter with an accurate and novel tuning architec-ture is presented. It exhibits 1EEE 802. 11a/b/g (9.5 MHz) and DVB-H (3 MHz, 4 MHz) application. The filter exploits digitally-controlled polysilicon resistor banks and a phase lock loop type automatic tuning system. The novel and complex automatic frequency calibration scheme provides better than 4 comer frequency accuracy, and it can be powered down after calibration to save power and avoid digital signal interference. The filter achieves OIP3 of 26 dBm and the measured group delay variation of the receiver filter is 50 ns (WLAN mode). Its dissipation is 3.4 mA in RX mode and 2.3 mA (only for one path) in TX mode from a 2.85 V supply. The dissipation of calibration consumes 2 mA. The circuit has been fabricated in a 0.35μm 47 GHz SiGe BiCMOS technology; the receiver and transmitter filter occupy 0.21 mm~2 and 0.11 mm~2 (calibration circuit excluded), respectively.

Investigation of transient process and steady output of lasing without inversion

The transient evolution processes and steady outputs of continuous wave lasing without inversion (LWI) and self-pulsing LWI in a resonant open V type three-level system are studied. It was found that the two kinds of LWI have some obvious differences not only from the steady outputs but also from the transient evolution processes. The effects of the unsaturated gain coefficient, cavity loss coefficient, ratio of the atomic injection rates and atomic exit rate on the transient evolution processes and steady outputs are discussed.

开放V型系统产生无反转激光的两类时间演化

从旋波、慢变振幅及平均场近似下的密度矩阵运动方程出发，利用数值计算结果研究了开放的V型三能级系统在共振条件下通过Pitchfork分岔产生的连续波无反转激光(LWI)场和通过Hopf分岔产生的自脉动LWI场及相应的各能级粒子数布居的时间演化规律，讨论了系统各参量变化对此时间演化规律的影响。

双暗态原子系统动力学行为的量子相干控制研究(英文)

量子相干控制前沿问题及应用研究是本世纪物理学前沿领域的重要研究内容.而基于暗态的量子相干控制技术已经导致了在相干布居捕获、绝热跟随、量子信息等多方面的应用.论文主要进行双暗态原子系统动力学行为的若干量子相干控制研究,包括双暗态四能级原子系统的绝热跟随特性研究,双暗态作用提高克尔非线性的新方案提出,自发辐射诱导相干实现非线性极化率的提高以及双通道高效四波混频过程的实现等.