A Trigger Enzyme in Mycoplasma pneumoniae: Impact of the Glycerophosphodiesterase GlpQ on Virulence and Gene Expression

Mycoplasma pneumoniae is a causative agent of atypical pneumonia. The formation of hydrogen peroxide, a product of glycerol metabolism, is essential for host cell cytotoxicity. Phosphatidylcholine is the major carbon source available on lung epithelia, and its utilization requires the cleavage of deacylated phospholipids to glycerol-3-phosphate and choline. M. pneumoniae possesses two potential glycerophosphodiesterases, MPN420 (GlpQ) and MPN566. In this work, the function of these proteins was analyzed by biochemical, genetic, and physiological studies. The results indicate that only GlpQ is an active glycerophosphodiesterase. MPN566 has no enzymatic activity as glycerophosphodiesterase and the inactivation of the gene did not result in any detectable phenotype. Inactivation of the glpQ gene resulted in reduced growth in medium with glucose as the carbon source, in loss of hydrogen peroxide production when phosphatidylcholine was present, and in a complete loss of cytotoxicity towards HeLa cells. All these phenotypes were reverted upon complementation of the mutant. Moreover, the glpQ mutant strain exhibited a reduced gliding velocity. A comparison of the proteomes of the wild type strain and the glpQ mutant revealed that this enzyme is also implicated in the control of gene expression. Several proteins were present in higher or lower amounts in the mutant. This apparent regulation by GlpQ is exerted at the level of transcription as determined by mRNA slot blot analyses. All genes subject to GlpQ-dependent control have a conserved potential cis-acting element upstream of the coding region. This element overlaps the promoter in the case of the genes that are repressed in a GlpQ-dependent manner and it is located upstream of the promoter for GlpQ-activated genes. We may suggest that GlpQ acts as a trigger enzyme that measures the availability of its product glycerol-3-phosphate and uses this information to differentially control gene expression

Imbalance of peptidoglycan biosynthesis alters the cell surface charge of Listeria monocytogenes

The bacterial cell wall is composed of a thick layer of peptidoglycan and cell wall polymers, which are either embedded in the membrane or linked to the peptidoglycan backbone and referred to as lipoteichoic acid (LTA) and wall teichoic acid (WTA), respectively. Modifications of the peptidoglycan or WTA backbone can alter the susceptibility of the bacterial cell towards cationic antimicrobials and lysozyme. The human pathogen Listeria monocytogenes is intrinsically resistant towards lysozyme, mainly due to deacetylation and O-acetylation of the peptidoglycan backbone via PgdA and OatA. Recent studies identified additional factors, which contribute to the lysozyme resistance of this pathogen. One of these is the predicted ABC transporter, EslABC. An eslB mutant is hyper-sensitive towards lysozyme, likely due to the production of thinner and less O-acetylated peptidoglycan. Using a suppressor screen, we show here that suppression of eslB phenotypes could be achieved by enhancing peptidoglycan biosynthesis, reducing peptidoglycan hydrolysis or alterations in WTA biosynthesis and modification. The lack of EslB also leads to a higher negative surface charge, which likely stimulates the activity of peptidoglycan hydrolases and lysozyme. Based on our results, we hypothesize that the portion of cell surface exposed WTA is increased in the eslB mutant due to the thinner peptidoglycan layer and that latter one could be caused by an impairment in UDP-N-acetylglucosamine (UDP-GlcNAc) production or distribution

Mycoplasma hyopneumoniae Transcription Unit Organization: Genome Survey and Prediction

Mycoplasma hyopneumoniae is associated with swine respiratory diseases. Although gene organization and regulation are well known in many prokaryotic organisms, knowledge on mycoplasma is limited. This study performed a comparative analysis of three strains of M. hyopneumoniae (7448, J and 232), with a focus on genome organization and gene comparison for open read frame (ORF) cluster (OC) identification. An in silico analysis of gene organization demonstrated 117 OCs and 34 single ORFs in M. hyopneumoniae 7448 and J, while 116 OCs and 36 single ORFs were identified in M. hyopneumoniae 232. Genomic comparison revealed high synteny and conservation of gene order between the OCs defined for 7448 and J strains as well as for 7448 and 232 strains. Twenty-one OCs were chosen and experimentally confirmed by reverse transcription–PCR from M. hyopneumoniae 7448 genome, validating our prediction. A subset of the ORFs within an OC could be independently transcribed due to the presence of internal promoters. Our results suggest that transcription occurs in ‘run-on’ from an upstream promoter in M. hyopneumoniae, thus forming large ORF clusters (from 2 to 29 ORFs in the same orientation) and indicating a complex transcriptional organization

Presupernova Evolution of Rotating Massive Stars I: Numerical Method and Evolution of the Internal Stellar Structure

The evolution of rotating stars with zero-age main sequence (ZAMS) masses in the range 8 to 25 M_sun is followed through all stages of stable evolution. The initial angular momentum is chosen such that the star's equatorial rotational velocity on the ZAMS ranges from zero to ~ 70 % of break-up. Redistribution of angular momentum and chemical species are then followed as a consequence of rotationally induced circulation and instablities. The effects of the centrifugal force on the stellar structure are included. Uncertain mixing efficiencies are gauged by observations. We find, as noted in previous work, that rotation increases the helium core masses and enriches the stellar envelopes with products of hydrogen burning. We determine, for the first time, the angular momentum distribution in typical presupernova stars along with their detailed chemical structure. Angular momentum loss due to (non-magnetic) stellar winds and the redistribution of angular momentum during core hydrogen burning are of crucial importance for the specific angular momentum of the core. Neglecting magnetic fields, we find angular momentum transport from the core to the envelope to be unimportant after core helium burning. We obtain specific angular momenta for the iron core and overlaying material of 1E16...1E17 erg s. These values are insensitive to the initial angular momentum. They are small enough to avoid triaxial deformations of the iron core before it collapses, but could lead to neutron stars which rotate close to break-up. They are also in the range required for the collapsar model of gamma-ray bursts. The apparent discrepancy with the measured rotation rates of young pulsars is discussed.Comment: 62 pages, including 7 tables and 19 figures. Accepted by Ap

Intermediate-velocity gas observed towards the Shajn 147 SNR

(Abridged) We present high-resolution spectra of the interstellar NaI and CaII absorption lines for 3 early-type stars along the line-of-sight towards the Shajn 147 SNR. These are supplemented with far-UV aborption spectra of HD 36665 and HD 37318 recorded with the NASA FUSE satellite. The observations reveal intermediate-velocity (IV) absorption features at V_helio = +92 km/s towards HD 37318 and -65 & -52 km/s towards HD 36665. These IV components can be associated with the expansion of the SNR. The observations suggest that the IV component at +92 km/s towards HD 37318 is composed mainly of warm and ionized gas. The two IV components observed towards HD 36665 are likely composed of both neutral and ionized gas shells. Highly ionized gas was detected in the OVI (1032\AA) absorption line at +40 km/s towards both stars. This hot gas component is characterized by a columnn density ratio (N(CIV)/N(OVI) < 0.27) consistent with that predicted by current models of evolved SNRs, but we cannot preclude its origin in the ISM. Column-density ratios have been derived for the IV gas components detected towards S147, as well as for fast-moving gas observed towards two other SNRs in order to gain some insight into the behavior of element abundances in the disturbed interstellar gas associated with these regions. In all cases except for Na and Ca, these elements appear to be present with near-solar abundance ratios. (Abridged)Comment: 11 pages, 5 figures, Latex, accepted for publication in A&

Be-star rotation: how close to critical?

We argue that, in general, observational studies of Be-star rotation have paid insufficient attention to the effects of equatorial gravity darkening. We present new line-profile calculations that emphasize the insensitivity of line width to rotation for fast rotators. Coupled with a critical review of observational procedures, these calculations suggest that the observational parameter v sin(i) may systematically underestimate the true projected equatorial rotation velocity, ve sin(i), by some tens of per cent for rapid rotators. The crucial implication of this work is that Be stars may be rotating much closer to their critical velocities than is generally supposed, bringing a range of new processes into contention for the elusive physical mechanism responsible for the circumstellar disk thought to be central to the Be phenomenon.Comment: 8 pages, 3 figures, shortened Sec. 6.3, new Sec. 6.4. Accepted by MNRA

Investigating the spectroscopic, magnetic and circumstellar variability of the O9 subgiant star HD 57682

The O9IV star HD 57682, discovered to be magnetic within the context of the MiMeS survey in 2009, is one of only eight convincingly detected magnetic O-type stars. Among this select group, it stands out due to its sharp-lined photospheric spectrum. Since its discovery, the MiMeS Collaboration has continued to obtain spectroscopic and magnetic observations in order to refine our knowledge of its magnetic field strength and geometry, rotational period, and spectral properties and variability. In this paper we report new ESPaDOnS spectropolarimetric observations of HD 57682, which are combined with previously published ESPaDOnS data and archival H{\alpha} spectroscopy. This dataset is used to determine the rotational period (63.5708 \pm 0.0057 d), refine the longitudinal magnetic field variation and magnetic geometry (dipole surface field strength of 880\pm50 G and magnetic obliquity of 79\pm4\circ as measured from the magnetic longitudinal field variations, assuming an inclination of 60\circ), and examine the phase variation of various lines. In particular, we demonstrate that the H{\alpha} equivalent width undergoes a double-wave variation during a single rotation of the star, consistent with the derived magnetic geometry. We group the variable lines into two classes: those that, like H{\alpha}, exhibit non-sinusoidal variability, often with multiple maxima during the rotation cycle, and those that vary essentially sinusoidally. Based on our modelling of the H{\alpha} emission, we show that the variability is consistent with emission being generated from an optically thick, flattened distribution of magnetically-confined plasma that is roughly distributed about the magnetic equator. Finally, we discuss our findings in the magnetospheric framework proposed in our earlier study.Comment: 21 pages, 19 figures, Accepted for publication in MNRA

Life on Arginine for Mycoplasma hominis: Clues from Its Minimal Genome and Comparison with Other Human Urogenital Mycoplasmas

Mycoplasma hominis is an opportunistic human mycoplasma. Two other pathogenic human species, M. genitalium and Ureaplasma parvum, reside within the same natural niche as M. hominis: the urogenital tract. These three species have overlapping, but distinct, pathogenic roles. They have minimal genomes and, thus, reduced metabolic capabilities characterized by distinct energy-generating pathways. Analysis of the M. hominis PG21 genome sequence revealed that it is the second smallest genome among self-replicating free living organisms (665,445 bp, 537 coding sequences (CDSs)). Five clusters of genes were predicted to have undergone horizontal gene transfer (HGT) between M. hominis and the phylogenetically distant U. parvum species. We reconstructed M. hominis metabolic pathways from the predicted genes, with particular emphasis on energy-generating pathways. The Embden–Meyerhoff–Parnas pathway was incomplete, with a single enzyme absent. We identified the three proteins constituting the arginine dihydrolase pathway. This pathway was found essential to promote growth in vivo. The predicted presence of dimethylarginine dimethylaminohydrolase suggested that arginine catabolism is more complex than initially described. This enzyme may have been acquired by HGT from non-mollicute bacteria. Comparison of the three minimal mollicute genomes showed that 247 CDSs were common to all three genomes, whereas 220 CDSs were specific to M. hominis, 172 CDSs were specific to M. genitalium, and 280 CDSs were specific to U. parvum. Within these species-specific genes, two major sets of genes could be identified: one including genes involved in various energy-generating pathways, depending on the energy source used (glucose, urea, or arginine) and another involved in cytadherence and virulence. Therefore, a minimal mycoplasma cell, not including cytadherence and virulence-related genes, could be envisaged containing a core genome (247 genes), plus a set of genes required for providing energy. For M. hominis, this set would include 247+9 genes, resulting in a theoretical minimal genome of 256 genes

Blue irregular variable stars in the Small Magellanic Cloud from EROS2 : Herbig Ae/Be or classical Be stars ?

Using data from the EROS2 microlensing survey, we report the discovery of two blue objects with irregular photometric behaviour of $\Delta V \sim 0.1$-0.4 mag on time scales of 20 to 200 days. They show a bluer when fainter behaviour. Subsequent spectra taken with the ESO 3.6m telescope show spectral type B4eIII and B2eIV-V with strong $H \alpha$ emission. These objects resemble the Herbig AeBe but also classical Be stars. At this stage, it is not possible to distinguish unambiguously between pre-main sequence and classical Be nature. If we favour the pre-main sequence interpretation, they are more luminous than the luminosity upper limit for Galactic HAeBe stars. The same was found for the HAeBe candidates in the LMC. This might be due to a shorter accretion time scale ($\tau = M_*/\dot{M}$), or the smaller dust content during the pre-main sequence evolution of SMC and LMC stars.Comment: : 9 pages, LaTeX, 7 figures. Accepted for publications in A