Yeast Polarity: Negative Feedback Shifts the Focus

A new study of Cdc42p polarization in yeast suggests that the actin cytoskeleton can destabilize the polarity axis, causing Cdc42p foci to wander aimlessly around the cell cortex

Mechanistic mathematical model of polarity in yeast

The establishment of cell polarity involves positive-feedback mechanisms that concentrate polarity regulators, including the conserved GTPase Cdc42p, at the “front” of the polarized cell. Previous studies in yeast suggested the presence of two parallel positive-feedback loops, one operating as a diffusion-based system, and the other involving actin-directed trafficking of Cdc42p on vesicles. F-actin (and hence directed vesicle traffic) speeds fluorescence recovery of Cdc42p after photobleaching, suggesting that vesicle traffic of Cdc42p contributes to polarization. We present a mathematical modeling framework that combines previously developed mechanistic reaction-diffusion and vesicle-trafficking models. Surprisingly, the combined model recapitulated the observed effect of vesicle traffic on Cdc42p dynamics even when the vesicles did not carry significant amounts of Cdc42p. Vesicle traffic reduced the concentration of Cdc42p at the front, so that fluorescence recovery mediated by Cdc42p flux from the cytoplasm took less time to replenish the bleached pool. Simulations in which Cdc42p was concentrated into vesicles or depleted from vesicles yielded almost identical predictions, because Cdc42p flux from the cytoplasm was dominant. These findings indicate that vesicle-mediated delivery of Cdc42p is not required to explain the observed Cdc42p dynamics, and raise the question of whether such Cdc42p traffic actually contributes to polarity establishment

Interaction between bud-site selection and polarity-establishment machineries in budding yeast

Saccharomyces cerevisiae yeast cells polarize in order to form a single bud in each cell cycle. Distinct patterns of bud-site selection are observed in haploid and diploid cells. Genetic approaches have identified the molecular machinery responsible for positioning the bud site: during bud formation, specific locations are marked with immobile landmark proteins. In the next cell cycle, landmarks act through the Ras-family GTPase Rsr1 to promote local activation of the conserved Rho-family GTPase, Cdc42. Additional Cdc42 accumulates by positive feedback, creating a concentrated patch of GTP-Cdc42, which polarizes the cytoskeleton to promote bud emergence. Using time-lapse imaging and mathematical modelling, we examined the process of bud-site establishment. Imaging reveals unexpected effects of the bud-site-selection system on the dynamics of polarity establishment, raising new questions about how that system may operate. We found that polarity factors sometimes accumulate at more than one site among the landmark-specified locations, and we suggest that competition between clusters of polarity factors determines the final location of the Cdc42 cluster. Modelling indicated that temporally constant landmark-localized Rsr1 would weaken or block competition, yielding more than one polarity site. Instead, we suggest that polarity factors recruit Rsr1, effectively sequestering it from other locations and thereby terminating landmark activity

Host-Bacteria Interactions in Foreign Body Infections

Persistent staphylococcal infections are a major medical problem, especially when they occur on implanted materials or intravascular catheters. This review describes some of the recently discovered molecular mechanisms of Staphylococcus aureus attachment to host proteins coating biomedical implants. These interactions involve specific surface proteins, called bacterial adhesins, that recognize specific domains of host proteins deposited on indwelling devices, such as fibronectin, fibrinogen, or fibrin. Elucidation of molecular mechanisms of S aureus adhesion to the different host proteins may lead to the development of specific inhibitors blocking attachment of S aureus, which may decrease the risk of bacterial colonization of indwelling device

Inhibition of Cdc42 during mitotic exit is required for cytokinesis

The role of Cdc42 and its regulation during cytokinesis is not well understood. Using biochemical and imaging approaches in budding yeast, we demonstrate that Cdc42 activation peaks during the G1/S transition and during anaphase but drops during mitotic exit and cytokinesis. Cdc5/Polo kinase is an important upstream cell cycle regulator that suppresses Cdc42 activity. Failure to down-regulate Cdc42 during mitotic exit impairs the normal localization of key cytokinesis regulators—Iqg1 and Inn1—at the division site, and results in an abnormal septum. The effects of Cdc42 hyperactivation are largely mediated by the Cdc42 effector p21-activated kinase Ste20. Inhibition of Cdc42 and related Rho guanosine triphosphatases may be a general feature of cytokinesis in eukaryotes

Cloud Atlas: Rotational Spectral Modulations and potential Sulfide Clouds in the Planetary-mass, Late T-type Companion Ross 458C

Measurements of photometric variability at different wavelengths provide insights into the vertical cloud structure of brown dwarfs and planetary-mass objects. In seven Hubble Space Telescope consecutive orbits, spanning $\sim$10 h of observing time}, we obtained time-resolved spectroscopy of the planetary-mass T8-dwarf Ross 458C using the near-infrared Wide Field Camera 3. We found spectrophotometric variability with a peak-to-peak signal of 2.62$\pm$0.02 % (in the 1.10-1.60~$\mu$m white light curve). Using three different methods, we estimated a rotational period of 6.75$\pm$1.58~h for the white light curve, and similar periods for narrow $J$- and $H$- band light curves. Sine wave fits to the narrow $J$- and $H$-band light curves suggest a tentative phase shift between the light curves with wavelength when we allow different periods between both light curves. If confirmed, this phase shift may be similar to the phase shift detected earlier for the T6.5 spectral type 2MASS J22282889-310262. We find that, in contrast with 2M2228, the variability of Ross~458C shows evidence for a {color trend} within the narrow $J$-band, but gray variations in the narrow $H$-band. The spectral time-resolved variability of Ross 458C might be potentially due to heterogeneous sulfide clouds in the atmosphere of the object. Our discovery extends the study of spectral modulations of condensate clouds to the coolest T dwarfs, planetary-mass companions.Comment: Accepted in ApJ

Comparative efficacies of imipenem, oxacillin and vancomycin for therapy of chronic foreign body infection due to methicillin-susceptible and -resistant Staphylococcus aureus

The efficacies of imipenem when directed against methicillin-susceptible (MSSA) and methicillin-resistant (MRSA) strains of Staphylococcus aureus were compared with those of oxacillin and vancomycin in a subcutaneous rat model, using chronically infected tissue cages. At three weeks after inoculation, stable chronic infections were established with average bacterial counts exceeding 106cfu/mL tissue cage fluid for both strains. Intraperitoneal administration (twice a day for 7 days) of imipenem (80 mg/kg) or oxacillin (200 mg/kg) produced peak levels of 23 or 45 mg/L and trough levels of < 0⋅1 and 5⋅7 mg/L, respectively. The therapeutic regimens of either imipenem (P < 0⋅001) or oxacillin (P < 0⋅02) administered for 7 days led to significant reductions in bacterial counts in the tissue cage fluids of animals chronically infected with MSSA. In contrast, imipenem was not effective against chronic MRSA tissue cage infections, despite the relatively low MIC of the infecting strain and the use of high dose (120 mg/kg) therapy. In-vitro susceptibility testings of MRSA performed before and after imipenem therapy demonstrated the emergence of a highly resistant subpopulatio

Cloud Atlas: Discovery of Rotational Spectral Modulations in a Low-mass, L-type Brown Dwarf Companion to a Star

Observations of rotational modulations of brown dwarfs and giant exoplanets allow the characterization of condensate cloud properties. As of now rotational spectral modulations have only been seen in three L-type brown dwarfs. We report here the discovery of rotational spectral modulations in LP261-75B, an L6-type intermediate surface gravity companion to an M4.5 star. As a part of the Cloud Atlas Treasury program we acquired time-resolved Wide Field Camera 3 grism spectroscopy (1.1--1.69~$\mu$m) of LP261-75B. We find gray spectral variations with the relative amplitude displaying only a weak wavelength dependence and no evidence for lower-amplitude modulations in the 1.4~$\mu$m water band than in the adjacent continuum. The likely rotational modulation period is 4.78$\pm$0.95 h, although the rotational phase is not well sampled. The minimum relative amplitude in the white light curve measured over the whole wavelength range is 2.41$\pm$0.14\%. We report an unusual light curve with seemingly three peaks approximately evenly distributed in rotational phase. The spectral modulations suggests that the upper atmosphere cloud properties in {LP261-75B} are similar to two other mid-L dwarfs of typical infrared colors, but differ from that of the extremely red L-dwarf WISE0047

The Mre11-Rad50-Nbs1 complex mediates activation of TopBP1 by ATM

The activation of ATR-ATRIP in response to double-stranded DNA breaks (DSBs) depends upon ATM in human cells and Xenopus egg extracts. One important aspect of this dependency involves regulation of TopBP1 by ATM. In Xenopus egg extracts, ATM associates with TopBP1 and thereupon phosphorylates it on S1131. This phosphorylation enhances the capacity of TopBP1 to activate the ATR-ATRIP complex. We show that TopBP1 also interacts with the Mre11-Rad50-Nbs1 (MRN) complex in egg extracts in a checkpoint-regulated manner. This interaction involves the Nbs1 subunit of the complex. ATM can no longer interact with TopBP1 in Nbs1-depleted egg extracts, which suggests that the MRN complex helps to bridge ATM and TopBP1 together. The association between TopBP1 and Nbs1 involves the first pair of BRCT repeats in TopBP1. In addition, the two tandem BRCT repeats of Nbs1 are required for this binding. Functional studies with mutated forms of TopBP1 and Nbs1 suggested that the BRCT-dependent association of these proteins is critical for a normal checkpoint response to DSBs. These findings suggest that the MRN complex is a crucial mediator in the process whereby ATM promotes the TopBP1-dependent activation of ATR-ATRIP in response to DSBs

Cloud Atlas: Rotational Modulations in the L/T Transition Brown Dwarf Companion HN Peg B

Time-resolved observations of brown dwarfs' rotational modulations provide powerful insights into the properties of condensate clouds in ultra-cool atmospheres. Multi-wavelength light curves reveal cloud vertical structures, condensate particle sizes, and cloud morphology, which directly constrain condensate cloud and atmospheric circulation models. We report results from Hubble Space Telescope/Wide Field Camera 3 (WFC3) near-infrared G141 taken in six consecutive orbits observations of HNPeg B, an L/T transition brown dwarf companion to a G0V type star. The best-fit sine wave to the 1.1 to 1.7 micron broadband light curve has the amplitude of and period of hour. The modulation amplitude has no detectable wavelength dependence except in the 1.4 micron water absorption band, indicating that the characteristic condensate particle sizes are large (greater than 1 micron). We detect significantly (4.4 sigma) lower modulation amplitude in the 1.4 micron water absorption band, and find that HN Peg B's spectral modulation resembles those of early T type brown dwarfs. We also describe a new empirical interpolation method to remove spectral contamination from the bright host star. This method may be applied in other high-contrast time-resolved observations with WFC3