Identification of the CysB-regulated gene, hslJ, related to the Escherichia coli novobiocin resistance phenotype

The cysB gene product is a LysR-type regulatory protein required for expression of the cys regulon. cysB mutants of Escherichia coli and Salmonella, along with being auxotrophs for the cysteine, exhibit increased resistance to the antibiotics novobiocin (Nov) and mecillinam. In this work, by using lambdaplacMu9 insertions creating random lacZ fusions, we identify a gene, hslJ, whose expression appeared to be increased in cysB mutants and needed for Nov resistance. Measurements of the hslJ::IacZ gene fusion expression demonstrated that the hslJ gene is negatively regulated by CysB. In addition we observe the negative autogenous control of HslJ. When the control imposed by CysB is lifted in the cysB mutant, the elevation of Nov resistance can be achieved only in the presence of wild-type hslJ allele. A double cysB hslJ mutant restores the sensitivity to Nov. Overexpression of the wild-type HslJ protein either in a cysB(+) or a cys(B-) background increases the level of Nov resistance indicating that hslJ product is indeed involved in accomplishing this phenotype. The hslJ::OmegaKan allele encodes the C-terminaly truncated mutant protein HslJ Q121Ter which is not functional in achieving the Nov resistance but when overexpressed induces the psp operon. Finally, we found that inactivation of hslJ does not affect the increased resistance to mecillinam in cysB mutants

The LysR-type transcriptional regulator CysB controls the repression of hslJ transcription in Escherichia coli

The LysR-type transcriptional regulator (LTTR) CysB is a transcription factor in Escherichia coli cells, where as a homotetramer it binds the target promoter regions and activates the genes involved in sulphur utilization and sulphonate-sulphur metabolism, while negatively autoregulating its own transcription. The hslJ gene was found to be negatively regulated by CysB and directly correlated with novobiocin resistance of the bacterium. cysB mutants showed upregulation of the hslJ:: lacZ gene fusion and exhibited increased novobiocin resistance. In this study the hslJ transcription start point and the corresponding putative sigma(70) promoter were determined. The hslJ promoter region was defined by employing different hslJ-lacZ operon fusions, and transcription of the hslJ gene was shown to be subject to both repression imposed by the CysB regulator and direct or indirect autogenous negative control. These two regulations compete to some extent but they are not mutually exclusive. CysB acts as a direct repressor of hslJ transcription and binds the hslJ promoter region that carries the putative CysB repressor site. This CysB binding, apparently responsible for repression, is enhanced in the presence of the ligand N-acetylserine (NAS), hitherto considered to be a positive cofactor in CysB-mediated gene regulations. Interallelic complementation of characterized CysB mutants I33N and S277Ter partially restored the repression of hslJ transcription and the consequent novobiocin sensitivity, but did not complement the cysteine auxotrophy

Spectroscopy and polarimetry of the gravitationally lensed quasar Q0957+561

We present new spectroscopic and polarimetric observations of the first discovered gravitational lens Q0957+561 obtained with the 6m telescope of the Special Astrophysical Observatory (SAO, Russia). We explore spectropolarimetric parameters of Q0957+561 A,B components to investigate the innermost structure of the quasar, and explore the nature of polarization in lensed quasars. Additionally, we compare their present-day spectral characteristics with previous observations in order to study long-term spectral changes. We analyze spectral characteristics of lensed quasar comparing spectra of A and B images, as well as comparing previously observed image spectra with present-day ones. The polarization parameters of A-B images are compared. We also model the macro-lens influence on the polarization of the images representing the gravitational lens with a singular isothermal elliptical potential. We find that the brightness and SED ratio of components A and B changed during a long period. Polarization in broad lines of components A and B showed that the equatorial scattering cannot be detected in this quasar. We find wavelength-dependent polarization that may be explained as a combination of the polarization from the disc and outflowing material. There is a significant difference between polarization parameters of the A and B images: the B component shows a higher polarization degree and polarization angle. However, both polarization vectors are nearly perpendicular to the observed radio jet projection. It indicates that the polarization in the continuum is coming from the accretion disc. Our simple lensing model of a polarized source showed that macro-lens can cause the observed differences in polarization parameters of Q0957+561 A,B images. Using Mg II broad line and luminosity of component A we estimated that the Q0957+561 black hole mass is M~(4.8-6.1) $10^8$ M$\odot$Comment: accepted in A

Spectroscopy and polarimetry of the gravitationally lensed quasar Q0957+561

Context. We present new spectroscopic and polarimetric observations of the first discovered gravitational lens, Q0957+561. The lensed quasar has been observed with the 6 m telescope of the Special Astrophysical Observatory (Russia) in polarimetric and spectroscopic modes. Aims. We explore the spectropolarimetric parameters of the A and B components of Q0957+561 to investigate the innermost structure of gravitationally lensed quasars and explore the nature of polarization in lensed quasars. Additionally, we aim to compare their present-day spectral characteristics with previous observations in order to study long-term spectral changes. Methods. We perform new spectral and polarization observations of the Q0957+561 A and B images. After observed data reduction, we analyse the spectral characteristics of the lensed quasar, comparing the spectra of the A and B images, as well as comparing previously observed image spectra with present-day ones. The polarization parameters of the two images are also compared. Furthermore, we model the macro-lens influence on the polarization of the images, representing the gravitational lens with a singular isothermal elliptical potential. Results. We find that the brightness and the spectral energy distribution ratio of components A and B have changed over a long period. Polarization in the broad lines of components A and B show that equatorial scattering cannot be detected in this lensed quasar. We find wavelength-dependent polarization that may be explained as a combination of the polarization from the disc and the outflowing material. There is a significant difference between the polarization parameters of the A and B images: The B component shows a higher polarization rate and polarization angle. However, both polarization vectors are nearly perpendicular to the observed radio jet projection. This indicates that the polarization in the continuum comes from the accretion disc. Our simple lensing model of a polarized source shows that, in principle, macro lenses can cause the observed differences in the polarization parameters of the Q0957+561A and B images. Using the Mg II broad line and luminosity of component A, we estimate the Q0957+561 black hole mass to be MSMBH ≈ (4.8 − 6.1) × 108 M⊙

Spectroscopy and polarimetry of the gravitationally lensed quasar SDSS J1004+4112 with the 6m SAO RAS telescope

Context. We present new spectroscopic and polarimetric observations of the gravitational lens SDSS J1004+4112 taken with the 6 m telescope of the Special Astrophysical Observatory (Russia). Aims. In order to explain the variability that is observed only in the blue wing of the C IV emission line, corresponding to image A, we analyze the spectroscopy and polarimetry of the four images of the lensed system. Methods. Spectra of the four images were taken in 2007, 2008, and 2018, and polarization was measured in the period 2014–2017. Additionally, we modeled the microlensing effect in the polarized light, assuming that the source of polarization is the equatorial scattering in the inner part of the torus. Results. We find that a blue enhancement in the C IV line wings affects component A in all three epochs. We also find that the UV continuum of component D was amplified in the period 2007–2008, and that the red wings of CIII] and C IV appear brighter in D than in the other three components. We report significant changes in the polarization parameters of image D, which can be explained by microlensing. Our simulations of microlensing of an equatorial scattering region in the dusty torus can qualitatively explain the observed changes in the polarization degree and angle of image D. We do not detect significant variability in the polarization parameters of the other images (A, B, and C), although the averaged values of the polarization degree and angle are different for the different images. Conclusions. Microlensing of a broad line region model including a compact outflowing component can qualitatively explain the C IV blue wing enhancement (and variation) in component A. However, to confirmed this hypothesis, we need additional spectroscopic observation in future