Identification and Characterization of Hells-CDCA7, a Nucleosome Remodeling Complex Required for Mitotic Chromosome Structure

The major components of mitotic chromosomes including histones, topoisomerase II and condensin are known to compact and shape the chromatin into rod-like chromatids. However, a complete picture of the proteins involved in shaping mitotic chromatin is unsettled. Here, I perform mass spectrometry on chromatin isolated from Xenopus egg extracts. By comparing interphase and M phase, I reveal how the chromatin proteome is affected by the cell cycle. I find that although topoisomerase II associates with chromatin throughout the cell cycle, its catalytic activity is greatly enhanced in mitosis where it can act on nucleosomal substrates. In contrast, condensin is specifically recruited to M phase chromatin and prefers a non-nucleosomal substrate. I show that nearly all proteins involved in nucleosome assembly and remodeling are evicted from mitotic chromosomes, concurrent with a reduction in mitotic nucleosome assembly and discuss a role for these phenomena in shaping mitotic chromosomes. In analyzing my mass spectrometry data, I noticed that subunits of stoichiometric chromatin-bound complexes behaved similarly across a variety of conditions. Using this principle, I identify a novel nucleosome remodeling complex comprising HELLS and CDCA7, two proteins known to be causative for Immunodeficiency, centromeric region instability, facial anomalies (ICF) syndrome, a rare immunodeficiency disease. Consistent with previous literature, HELLS alone fails to remodel nucleosomes, but the HELLS-CDCA7 complex possesses robust nucleosome remodeling activity. CDCA7 is essential for loading HELLS onto chromatin, and CDCA7 harboring patient ICF mutations fails to recruit the complex to chromatin. Finally, I show that the HELLSCDCA7 complex is required for proper mitotic chromosome structure. Together, my study identifies a unique bipartite nucleosome remodeling complex where the functional remodeling activity is split between two proteins. These data lead me to a unifying model for ICF syndrome where all known ICF mutations converge on defective DNA methylation. I propose that HELLS-CDCA7 mediated remodeling of juxtacentromere heterochromatic nucleosomes facilitates DNMT3B-mediated methylation and discuss this model alongside alternative models for ICF syndrome. Finally, I consider the role of CDCA7 paralogs in differentially regulating HELLS, and present an outlook on the future of ICF syndrome research

A near IR imaging survey of intermediate and high-mass young stellar outflow candidates

We have carried out a near-infrared imaging survey of luminous young stellar outflow candidates using the United Kingdom Infrared Telescope. Observations were obtained in the broad band K (2.2 mu) and through narrow band filters at the wavelengths of H_2 v=1--0 S(1) (2.1218 mu) and Br gamma (2.166 mu) lines. Fifty regions were imaged with a field of view of 2.2 X 2.2 arcmin^2. Several young embedded clusters are unveiled in our near-infrared images. 76% of the objects exhibit H_2 emission and 50% or more of the objects exhibit aligned H_2 emission features suggesting collimated outflows, many of which are new detections. These observations suggest that disk accretion is probably the leading mechanism in the formation of stars, at least up to late O spectral types. The young stellar objects responsible for many of these outflows are positively identified in our images based on their locations with respect to the outflow lobes, 2MASS colours and association with MSX, IRAS, millimetre and radio sources. The close association of molecular outflows detected in CO with the H_2 emission features produced by shock excitation by jets from the young stellar objects suggests that the outflows from these objects are jet-driven. Towards strong radio emitting sources, H_2 jets were either not detected or were weak when detected, implying that most of the accretion happens in the pre-UCHII phase; accretion and outflows are probably weak when the YSO has advanced to its UCHII stage.Comment: 64 pages, 53 figures, Accepted for publication in the MNRA

LSST: from Science Drivers to Reference Design and Anticipated Data Products

(Abridged) We describe here the most ambitious survey currently planned in the optical, the Large Synoptic Survey Telescope (LSST). A vast array of science will be enabled by a single wide-deep-fast sky survey, and LSST will have unique survey capability in the faint time domain. The LSST design is driven by four main science themes: probing dark energy and dark matter, taking an inventory of the Solar System, exploring the transient optical sky, and mapping the Milky Way. LSST will be a wide-field ground-based system sited at Cerro Pach\'{o}n in northern Chile. The telescope will have an 8.4 m (6.5 m effective) primary mirror, a 9.6 deg$^2$ field of view, and a 3.2 Gigapixel camera. The standard observing sequence will consist of pairs of 15-second exposures in a given field, with two such visits in each pointing in a given night. With these repeats, the LSST system is capable of imaging about 10,000 square degrees of sky in a single filter in three nights. The typical 5$\sigma$ point-source depth in a single visit in $r$ will be $\sim 24.5$ (AB). The project is in the construction phase and will begin regular survey operations by 2022. The survey area will be contained within 30,000 deg$^2$ with $\delta<+34.5^\circ$, and will be imaged multiple times in six bands, $ugrizy$, covering the wavelength range 320--1050 nm. About 90\% of the observing time will be devoted to a deep-wide-fast survey mode which will uniformly observe a 18,000 deg$^2$ region about 800 times (summed over all six bands) during the anticipated 10 years of operations, and yield a coadded map to $r\sim27.5$. The remaining 10\% of the observing time will be allocated to projects such as a Very Deep and Fast time domain survey. The goal is to make LSST data products, including a relational database of about 32 trillion observations of 40 billion objects, available to the public and scientists around the world.Comment: 57 pages, 32 color figures, version with high-resolution figures available from https://www.lsst.org/overvie

LSST control software component design

Construction of the Large Synoptic Survey Telescope system involves several different organizations, a situation that poses many challenges at the time of the software integration of the components. To ensure commonality for the purposes of usability, maintainability, and robustness, the LSST software teams have agreed to the following for system software components: a summary state machine, a manner of managing settings, a flexible solution to specify controller/controllee relationships reliably as needed, and a paradigm for responding to and communicating alarms. This paper describes these agreed solutions and the factors that motivated these

The Spitzer survey of interstellar clouds in the Gould Belt. II. The Cepheus Flare observed with IRAC and MIPS

We present Spitzer IRAC (~2 deg^2) and MIPS (~8 deg^2) observations of the Cepheus Flare which is associated with the Gould Belt, at an approximate distance of ~300 pc. Around 6500 sources are detected in all four IRAC bands, of which ~900 have MIPS 24 micron detections. We identify 133 YSO candidates using color-magnitude diagram techniques, a large number of the YSO candidates are associated with the NGC 7023 reflection nebula. Cross identifications were made with the Guide Star Catalog II and the IRAS Faint Source Catalog, and spectral energy distributions (SED) were constructed. SED modeling was conducted to estimate the degree of infrared excess. It was found that a large majority of disks were optically thick accreting disks, suggesting that there has been little disk evolution in these sources. Nearest-neighbor clustering analysis identified four small protostellar groups (L1228, L1228N, L1251A, and L1251B) with 5-15 members each and the larger NGC 7023 association with 32 YSO members. The star formation efficiency for cores with clusters of protostars and for those without clusters was found to be ~8% and ~1% respectively. The cores L1155, L1241, and L1247 are confirmed to be starless down to our luminosity limit of L_bol=0.06 L_sol.Comment: Submitted to ApJSS. 20 figures, 110 page

Energetic eruptions leading to a peculiar hydrogen-rich explosion of a massive star

Every supernova so far observed has been considered to be the terminal explosion of a star. Moreover, all supernovae with absorption lines in their spectra show those lines decreasing in velocity over time, as the ejecta expand and thin, revealing slower-moving material that was previously hidden. In addition, every supernova that exhibits the absorption lines of hydrogen has one main light-curve peak, or a plateau in luminosity, lasting approximately 100 days before declining1. Here we report observations of iPTF14hls, an event that has spectra identical to a hydrogen-rich core-collapse supernova, but characteristics that differ extensively from those of known supernovae. The light curve has at least five peaks and remains bright for more than 600 days; the absorption lines show little to no decrease in velocity; and the radius of the line-forming region is more than an order of magnitude bigger than the radius of the photosphere derived from the continuum emission. These characteristics are consistent with a shell of several tens of solar masses ejected by the progenitor star at supernova-level energies a few hundred days before a terminal explosion. Another possible eruption was recorded at the same position in 1954. Multiple energetic pre-supernova eruptions are expected to occur in stars of 95 to 130 solar masses, which experience the pulsational pair instability2,3,4,5. That model, however, does not account for the continued presence of hydrogen, or the energetics observed here. Another mechanism for the violent ejection of mass in massive stars may be required

Yeast Mutants Affecting Possible Quality Control of Plasma Membrane Proteins

Mutations gef1, stp22, STP26, and STP27 in Saccharomyces cerevisiae were identified as suppressors of the temperature-sensitive α-factor receptor (mutation ste2-3) and arginine permease (mutation can1(ts)). These suppressors inhibited the elimination of misfolded receptors (synthesized at 34°C) as well as damaged surface receptors (shifted from 22 to 34°C). The stp22 mutation (allelic to vps23 [M. Babst and S. Emr, personal communication] and the STP26 mutation also caused missorting of carboxypeptidase Y, and ste2-3 was suppressed by mutations vps1, vps8, vps10, and vps28 but not by mutation vps3. In the stp22 mutant, both the mutant and the wild-type receptors (tagged with green fluorescent protein [GFP]) accumulated within an endosome-like compartment and were excluded from the vacuole. GFP-tagged Stp22p also accumulated in this compartment. Upon reaching the vacuole, cytoplasmic domains of both mutant and wild-type receptors appeared within the vacuolar lumen. Stp22p and Gef1p are similar to tumor susceptibility protein TSG101 and voltage-gated chloride channel, respectively. These results identify potential elements of plasma membrane quality control and indicate that cytoplasmic domains of membrane proteins are translocated into the vacuolar lumen