Genetic analysis of the spindle checkpoint genes san-1, mdf-2, bub-3 and the CENP-F homologues hcp-1 and hcp-2 in Caenorhabditis elegans

<p>Abstract</p> <p>Background</p> <p>The spindle checkpoint delays the onset of anaphase until all sister chromatids are aligned properly at the metaphase plate. To investigate the role <it>san-1</it>, the MAD3 homologue, has in <it>Caenorhabditis elegans </it>embryos we used RNA interference (RNAi) to identify genes synthetic lethal with the viable <it>san-1(ok1580) </it>deletion mutant.</p> <p>Results</p> <p>The <it>san-1(ok1580) </it>animal has low penetrating phenotypes including an increased incidence of males, larvae arrest, slow growth, protruding vulva, and defects in vulva morphogenesis. We found that the viability of <it>san-1(ok1580) </it>embryos is significantly reduced when HCP-1 (CENP-F homologue), MDF-1 (MAD-1 homologue), MDF-2 (MAD-2 homologue) or BUB-3 (predicted BUB-3 homologue) are reduced by RNAi. Interestingly, the viability of <it>san-1(ok1580) </it>embryos is not significantly reduced when the paralog of HCP-1, HCP-2, is reduced. The phenotype of <it>san-1(ok1580);hcp-1(RNAi) </it>embryos includes embryonic and larval lethality, abnormal organ development, and an increase in abnormal chromosome segregation (aberrant mitotic nuclei, anaphase bridging). Several of the <it>san-1(ok1580);hcp-1(RNAi) </it>animals displayed abnormal kinetochore (detected by MPM-2) and microtubule structure. The survival of <it>mdf-2(RNAi);hcp-1(RNAi) </it>embryos but not <it>bub-3(RNAi);hcp-1(RNAi) </it>embryos was also compromised. Finally, we found that <it>san-1(ok1580) </it>and <it>bub-3(RNAi)</it>, but not <it>hcp-1(RNAi) </it>embryos, were sensitive to anoxia, suggesting that like SAN-1, BUB-3 has a functional role as a spindle checkpoint protein.</p> <p>Conclusion</p> <p>Together, these data suggest that in the <it>C. elegans </it>embryo, HCP-1 interacts with a subset of the spindle checkpoint pathway. Furthermore, the fact that <it>san-1(ok1580);hcp-1(RNAi) </it>animals had a severe viability defect whereas in the <it>san-1(ok1580);hcp-2(RNAi) </it>and <it>san-1(ok1580);hcp-2(ok1757) </it>animals the viability defect was not as severe suggesting that <it>hcp-1 </it>and <it>hcp-2 </it>are not completely redundant.</p

The M81 Group Dwarf Irregular Galaxy DDO 165. I. High Velocity Neutral Gas in a Post-Starburst System

We present new multi-configuration VLA HI spectral line observations of the M81 group dIrr post-starburst galaxy DDO 165. The HI morphology is complex, with multiple column density peaks surrounding a large region of very low HI surface density that is offset from the center of the stellar distribution. The bulk of the neutral gas is associated with the southern section of the galaxy; a secondary peak in the north contains ~15% of the total HI mass. These components appear to be kinematically distinct, suggesting that either tidal processes or large-scale blowout have recently shaped the ISM of DDO 165. Using spatially-resolved position-velocity maps, we find multiple localized high-velocity gas features. Cross-correlating with radius-velocity analyses, we identify eight shell/hole structures in the ISM with a range of sizes (~400-900 pc) and expansion velocities (~7-11 km/s). These structures are compared with narrow- and broad-band imaging from KPNO and HST. Using the latter data, recent works have shown that DDO 165's previous "burst" phase was extended temporally (>1 Gyr). We thus interpret the high-velocity gas features, HI holes, and kinematically distinct components of the galaxy in the context of the immediate effects of "feedback" from recent star formation. In addition to creating HI holes and shells, extended star formation events are capable of creating localized high velocity motion of the surrounding interstellar material. A companion paper connects the energetics from the HI and HST data.Comment: The Astrophysical Journal, in press. Full-resolution version available on request from the first autho

Mycobacterium tuberculosis ClpP Proteases Are Co-transcribed but Exhibit Different Substrate Specificities

PMCID: PMC3613350This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

The ACS Nearby Galaxy Survey Treasury II. Young Stars and their Relation to Halpha and UV Emission Timescales in the M81 Outer Disk

We have obtained resolved stellar photometry from Hubble Space Telescope (HST) Advanced Camera for Surveys (ACS) observations of a field in the outer disk of M81 as part of the ACS Nearby Galaxy Survey Treasury (ANGST). Motivated by the recent discovery of extended UV (XUV) disks around many nearby spiral galaxies, we use the observed stellar population to derive the recent star formation histories of five ~0.5 kpc-sized regions within this field. These regions were selected on the basis of their UV luminosity from GALEX and include two HII regions, two regions which are UV-bright but Halpha-faint, and one "control" region faint in both UV and Halpha. We estimate our effective SFR detection limit at ~2 x 10^-4 Msun/yr, which is lower than that of GALEX for regions of this size. As expected, the HII regions contain massive main sequence stars (in the mass range 18-27 Msun, based on our best extinction estimates), while similar massive main sequence stars are lacking in the UV-bright/Halpha-faint regions. The observations are consistent with stellar ages 16 Myr in the UV-bright/Halpha-faint regions. All regions but the control have formed ~10^4 Msun of stars over the past ~65 Myr. Thus, our results, for at least one small area in the outer disk of M81, are consistent with an age difference being sufficient to explain the observed discrepancy between star-forming regions detected in Halpha and those detected exclusively in UV. However, our data cannot conclusively rule out other explanations, such as a strongly truncated initial mass function (IMF).Comment: 18 pages, 12 figures, accepted for publication in ApJ, paper with full resolution figures available: http://www.nearbygalaxies.org/papers/M81_Halpha_uv.pd

Using Drugs to Probe the Variability of Trans-Epithelial Airway Resistance

BACKGROUND:Precision medicine aims to combat the variability of the therapeutic response to a given medicine by delivering the right medicine to the right patient. However, the application of precision medicine is predicated on a prior quantitation of the variance of the reference range of normality. Airway pathophysiology provides a good example due to a very variable first line of defence against airborne assault. Humans differ in their susceptibility to inhaled pollutants and pathogens in part due to the magnitude of trans-epithelial resistance that determines the degree of epithelial penetration to the submucosal space. This initial 'set-point' may drive a sentinel event in airway disease pathogenesis. Epithelia differentiated in vitro from airway biopsies are commonly used to model trans-epithelial resistance but the 'reference range of normality' remains problematic. We investigated the range of electrophysiological characteristics of human airway epithelia grown at air-liquid interface in vitro from healthy volunteers focusing on the inter- and intra-subject variability both at baseline and after sequential exposure to drugs modulating ion transport. METHODOLOGY/PRINCIPAL FINDINGS:Brushed nasal airway epithelial cells were differentiated at air-liquid interface generating 137 pseudostratified ciliated epithelia from 18 donors. A positively-skewed baseline range exists for trans-epithelial resistance (Min/Max: 309/2963 Ω·cm2), trans-epithelial voltage (-62.3/-1.8 mV) and calculated equivalent current (-125.0/-3.2 μA/cm2; all non-normal, P<0.001). A minority of healthy humans manifest a dramatic amiloride sensitivity to voltage and trans-epithelial resistance that is further discriminated by prior modulation of cAMP-stimulated chloride transport. CONCLUSIONS/SIGNIFICANCE:Healthy epithelia show log-order differences in their ion transport characteristics, likely reflective of their initial set-points of basal trans-epithelial resistance and sodium transport. Our data may guide the choice of the background set point in subjects with airway diseases and frame the reference range for the future delivery of precision airway medicine

Pathogenic variants in SLF2 and SMC5 cause segmented chromosomes and mosaic variegated hyperploidy

Embryonic development is dictated by tight regulation of DNA replication, cell division and differentiation. Mutations in DNA repair and replication genes disrupt this equilibrium, giving rise to neurodevelopmental disease characterized by microcephaly, short stature and chromosomal breakage. Here, we identify biallelic variants in two components of the RAD18-SLF1/2-SMC5/6 genome stability pathway, SLF2 and SMC5, in 11 patients with microcephaly, short stature, cardiac abnormalities and anemia. Patient-derived cells exhibit a unique chromosomal instability phenotype consisting of segmented and dicentric chromosomes with mosaic variegated hyperploidy. To signify the importance of these segmented chromosomes, we have named this disorder Atelís (meaning - incomplete) Syndrome. Analysis of Atelís Syndrome cells reveals elevated levels of replication stress, partly due to a reduced ability to replicate through G-quadruplex DNA structures, and also loss of sister chromatid cohesion. Together, these data strengthen the functional link between SLF2 and the SMC5/6 complex, highlighting a distinct role for this pathway in maintaining genome stability

Understanding Communication Signals during Mycobacterial Latency through Predicted Genome-Wide Protein Interactions and Boolean Modeling

About 90% of the people infected with Mycobacterium tuberculosis carry latent bacteria that are believed to get activated upon immune suppression. One of the fundamental challenges in the control of tuberculosis is therefore to understand molecular mechanisms involved in the onset of latency and/or reactivation. We have attempted to address this problem at the systems level by a combination of predicted functional protein∶protein interactions, integration of functional interactions with large scale gene expression studies, predicted transcription regulatory network and finally simulations with a Boolean model of the network. Initially a prediction for genome-wide protein functional linkages was obtained based on genome-context methods using a Support Vector Machine. This set of protein functional linkages along with gene expression data of the available models of latency was employed to identify proteins involved in mediating switch signals during dormancy. We show that genes that are up and down regulated during dormancy are not only coordinately regulated under dormancy-like conditions but also under a variety of other experimental conditions. Their synchronized regulation indicates that they form a tightly regulated gene cluster and might form a latency-regulon. Conservation of these genes across bacterial species suggests a unique evolutionary history that might be associated with M. tuberculosis dormancy. Finally, simulations with a Boolean model based on the regulatory network with logical relationships derived from gene expression data reveals a bistable switch suggesting alternating latent and actively growing states. Our analysis based on the interaction network therefore reveals a potential model of M. tuberculosis latency