Genomic Adaptations to the Loss of a Conserved Bacterial DNA Methyltransferase.

UNLABELLED: CcrM is an orphan DNA methyltransferase nearly universally conserved in a vast group of Alphaproteobacteria. In Caulobacter crescentus, it controls the expression of key genes involved in the regulation of the cell cycle and cell division. Here, we demonstrate, using an experimental evolution approach, that C. crescentus can significantly compensate, through easily accessible genetic changes like point mutations, the severe loss in fitness due to the absence of CcrM, quickly improving its growth rate and cell morphology in rich medium. By analyzing the compensatory mutations genome-wide in 12 clones sampled from independent ΔccrM populations evolved for ~300 generations, we demonstrated that each of the twelve clones carried at least one mutation that potentially stimulated ftsZ expression, suggesting that the low intracellular levels of FtsZ are the major burden of ΔccrM mutants. In addition, we demonstrate that the phosphoenolpyruvate-carbohydrate phosphotransfer system (PTS) actually modulates ftsZ and mipZ transcription, uncovering a previously unsuspected link between metabolic regulation and cell division in Alphaproteobacteria. We present evidence that point mutations found in genes encoding proteins of the PTS provide the strongest fitness advantage to ΔccrM cells cultivated in rich medium despite being disadvantageous in minimal medium. This environmental sign epistasis might prevent such mutations from getting fixed under changing natural conditions, adding a plausible explanation for the broad conservation of CcrM. IMPORTANCE: In bacteria, DNA methylation has a variety of functions, including the control of DNA replication and/or gene expression. The cell cycle-regulated DNA methyltransferase CcrM modulates the transcription of many genes and is critical for fitness in Caulobacter crescentus. Here, we used an original experimental evolution approach to determine which of its many targets make CcrM so important physiologically. We show that populations lacking CcrM evolve quickly, accumulating an excess of mutations affecting, directly or indirectly, the expression of the ftsZ cell division gene. This finding suggests that the most critical function of CcrM in C. crescentus is to promote cell division by enhancing FtsZ intracellular levels. During this work, we also discovered an unexpected link between metabolic regulation and cell division that might extend to other Alphaproteobacteria

Effects of (p)ppGpp on the progression of the cell cycle of Caulobacter crescentus.

Bacteria must control the progression of their cell cycle in response to nutrient availability. This regulation can be mediated by guanosine tetra- or pentaphosphate [(p)ppGpp], which are synthesized by enzymes of the RelA/SpoT homologue (Rsh) family, particularly under starvation conditions. Here, we study the effects of (p)ppGpp on the cell cycle of Caulobacter crescentus, an oligotrophic bacterium with a dimorphic life cycle. C. crescentus divides asymmetrically, producing a motile swarmer cell that cannot replicate its chromosome and a sessile stalked cell that is replication competent. The swarmer cell rapidly differentiates into a stalked cell in appropriate conditions. An artificial increase in the levels of (p)ppGpp in nonstarved C. crescentus cells was achieved by expressing a truncated relA gene from Escherichia coli, encoding a constitutively active (p)ppGpp synthetase. By combining single-cell microscopy, flow cytometry approaches, and swarming assays, we show that an increase in the intracellular concentration of (p)ppGpp is sufficient to slow down the swarmer-to-stalked cell differentiation process and to delay the initiation of chromosome replication. We also present evidence that the intracellular levels of two master regulators of the cell cycle of C. crescentus, DnaA and CtrA, are modulated in response to (p)ppGpp accumulation, even in the absence of actual starvation. CtrA proteolysis and DnaA synthesis seem indirectly inhibited by (p)ppGpp accumulation. By extending the life span of the motile nonreproductive swarmer cell and thus promoting dispersal and foraging functions over multiplication under starvation conditions, (p)ppGpp may play a central role in the ecological adaptation of C. crescentus to nutritional stresses

Enhanced heat transport by turbulent two-phase Rayleigh-B\'enard convection

We report measurements of turbulent heat-transport in samples of ethane (C$_2$H$_6$) heated from below while the applied temperature difference $\Delta T$ straddled the liquid-vapor co-existance curve $T_\phi(P)$. When the sample top temperature $T_t$ decreased below $T_\phi$, droplet condensation occurred and the latent heat of vaporization $H$ provided an additional heat-transport mechanism.The effective conductivity $\lambda_{eff}$ increased linearly with decreasing $T_t$, and reached a maximum value $\lambda_{eff}^*$ that was an order of magnitude larger than the single-phase $\lambda_{eff}$. As $P$ approached the critical pressure, $\lambda_{eff}^*$ increased dramatically even though $H$ vanished. We attribute this phenomenon to an enhanced droplet-nucleation rate as the critical point is approached.Comment: 4 gages, 6 figure

Magnetic Structure of Rapidly Rotating FK Comae-Type Coronae

We present a three-dimensional simulation of the corona of an FK Com-type rapidly rotating G giant using a magnetohydrodynamic model that was originally developed for the solar corona in order to capture the more realistic, non-potential coronal structure. We drive the simulation with surface maps for the radial magnetic field obtained from a stellar dynamo model of the FK Com system. This enables us to obtain the coronal structure for different field topologies representing different periods of time. We find that the corona of such an FK Com-like star, including the large scale coronal loops, is dominated by a strong toroidal component of the magnetic field. This is a result of part of the field being dragged by the radial outflow, while the other part remains attached to the rapidly rotating stellar surface. This tangling of the magnetic field,in addition to a reduction in the radial flow component, leads to a flattening of the gas density profile with distance in the inner part of the corona. The three-dimensional simulation provides a global view of the coronal structure. Some aspects of the results, such as the toroidal wrapping of the magnetic field, should also be applicable to coronae on fast rotators in general, which our study shows can be considerably different from the well-studied and well-observed solar corona. Studying the global structure of such coronae should also lead to a better understanding of their related stellar processes, such as flares and coronal mass ejections, and in particular, should lead to an improved understanding of mass and angular momentum loss from such systems.Comment: Accepted to ApJ, 10 pages, 6 figure

Instrumentation for hydrogen slush storage containers

Hydrogen liquid and slush tank continuous inventory during ground storag

Surface differential rotation and prominences of the Lupus post T Tauri star RX J1508.6-4423

We present in this paper a spectroscopic monitoring of the Lupus post T Tauri star RX J1508.6-4423 carried out at two closely separated epochs (1998 May 06 and 10) with the UCL Echelle Spectrograph on the 3.9-m Anglo-Australian Telescope. Applying least-squares convolution and maximum entropy image reconstruction techniques to our sets of spectra, we demonstrate that this star features on its surface a large cool polar cap with several appendages extending to lower latitudes, as well as one spot close to the equator. The images reconstructed at both epochs are in good overall agreement, except for a photospheric shear that we interpret in terms of latitudinal differential rotation. Given the spot distribution at the epoch of our observations, differential rotation could only be investigated between latitudes 15° and 60°. We find in particular that the observed differential rotation is compatible with a solar-like law (i.e., with rotation rate decreasing towards high latitudes proportionally to sin 2l, where l denotes the latitude) in this particular latitude range. Assuming that such a law can be extrapolated to all latitudes, we find that the equator of RX J1508.6-4423 does one more rotational cycle than the pole every 50 ±10 d, implying a photospheric shear 2 to 3 times stronger than that of the Sun. We also discover that the Hα emission profile of RX J1508.6-4423 is most of the time double-peaked and strongly modulated with the rotation period of the star. We interpret this rotationally modulated emission as being caused by a dense and complex prominence system, the circumstellar distribution of which is obtained through maximum entropy Doppler tomography. These maps show in particular that prominences form a complete and inhomogeneous ring around the star, precisely at the corotation radius. We use the total Hα and Hβ emission flux to estimate that the mass of the whole prominence system is about 10 20g. From our observation that the whole cloud system surrounding the star is regenerated in less than 4 d, we conclude that the braking time-scale of RX J1508.6-4423 is shorter than 1 Gyr, and that prominence expulsion is thus likely to contribute significantly to the rotational spindown of young low-mass stars

A search for starlight reflected from HD 75289 b

We have used a doppler tomographic analysis to conduct a deep search for the starlight reflected from the planetary companion to HD 75289. In 4 nights on VLT2/UVES in January 2003, we obtained 684 high resolution echelle spectra with a total integration time of 26 hours. We establish an upper limit on the planet's geometric albedo p < 0.12 (to the 99.9% significance level) at the most probable orbital inclination i ~ 60 degrees, assuming a grey albedo, a Venus-like phase function and a planetary radius R_p = 1.6 R_Jup. We are able to rule out some combinations of the predicted planetary radius and atmospheric albedo models with high, reflective cloud decks.Comment: 5 pages, 5 figures, MNRAS accepted 12 Oct 200