Tracking autophagy during proliferation and differentiation of trypanosoma brucei

Autophagy is a lysosome-dependent degradation mechanism that sequesters target cargo into autophagosomal vesicles. The Trypanosoma brucei genome contains apparent orthologues of several autophagy-related proteins including an ATG8 family. These ubiquitin-like proteins are required for autophagosome membrane formation, but our studies show that ATG8.3 is atypical. To investigate the function of other ATG proteins, RNAi compatible T. brucei were modified to function as autophagy reporter lines by expressing only either YFP-ATG8.1 or YFP-ATG8.2. In the insect procyclic lifecycle stage, independent RNAi down-regulation of ATG3 or ATG7 generated autophagy-defective mutants and confirmed a pro-survival role for autophagy in the procyclic form nutrient starvation response. Similarly, RNAi depletion of ATG5 or ATG7 in the bloodstream form disrupted autophagy, but did not impede proliferation. Further characterisation showed bloodstream form autophagy mutants retain the capacity to undergo the complex cellular remodelling that occurs during differentiation to the procyclic form and are equally susceptible to dihydroxyacetone-induced cell death as wild type parasites, not supporting a role for autophagy in this cell death mechanism. The RNAi reporter system developed, which also identified TOR1 as a negative regulator controlling YFP-ATG8.2 but not YFP-ATG8.1 autophagosome formation, will enable further targeted analysis of the mechanisms and function of autophagy in the medically relevant bloodstream form of T. brucei

ALMA CO J=6-5 observations of IRAS16293-2422: Shocks and entrainment

Observations of higher-excited transitions of abundant molecules such as CO are important for determining where energy in the form of shocks is fed back into the parental envelope of forming stars. The nearby prototypical and protobinary low-mass hot core, IRAS16293-2422 (I16293) is ideal for such a study. The source was targeted with ALMA for science verification purposes in band 9, which includes CO J=6-5 (E_up/k_B ~ 116 K), at an unprecedented spatial resolution (~0.2", 25 AU). I16293 itself is composed of two sources, A and B, with a projected distance of 5". CO J=6-5 emission is detected throughout the region, particularly in small, arcsecond-sized hotspots, where the outflow interacts with the envelope. The observations only recover a fraction of the emission in the line wings when compared to data from single-dish telescopes, with a higher fraction of emission recovered at higher velocities. The very high angular resolution of these new data reveal that a bow shock from source A coincides, in the plane of the sky, with the position of source B. Source B, on the other hand, does not show current outflow activity. In this region, outflow entrainment takes place over large spatial scales, >~ 100 AU, and in small discrete knots. This unique dataset shows that the combination of a high-temperature tracer (e.g., CO J=6-5) and very high angular resolution observations is crucial for interpreting the structure of the warm inner environment of low-mass protostars.Comment: Accepted for publication in A&A Letter

The RMS Survey: Resolving kinematic distance ambiguities towards a sample of compact HII regions using HI absorption

We present high-resolution HI data obtained using the Australia Telescope Compact Array to resolve the near/far distance ambiguities towards a sample of compact HII regions from the Red MSX Source (RMS) survey. The high resolution data are complemented with lower resolution archival HI data extracted from the Southern and VLA Galactic Plane surveys. We resolve the distance ambiguity for nearly all of the 105 sources where the continuum was strong enough to allow analysis of the HI absorption line structure. This represents another step in the determination of distances to the total RMS sample, which with over 1,000 massive young stellar objects and compact HII regions, is the largest and most complete sample of its kind. The full sample will allow the distribution of massive star formation in the Galaxy to be examined.Comment: Accepted by MNRAS. This paper consists of 15 pages and contains 10 figures and 5 table

VLTI/MIDI 10 micron interferometry of the forming massive star W33A

We report on resolved interferometric observations with VLTI/MIDI of the massive young stellar object (MYSO) W33A. The MIDI observations deliver spectrally dispersed visibilities with values between 0.03 and 0.06, for a baseline of 45m over the wavelength range 8-13 micron. The visibilities indicate that W33A has a FWHM size of approximately 120AU (0.030'') at 8 micron which increases to 240AU at 13 micron, scales previously unexplored among MYSOs. This observed trend is consistent with the temperature falling off with distance. 1D dust radiative transfer models are simultaneously fit to the visibility spectrum, the strong silicate feature and the shape of the mid infrared spectral energy distribution (SED). For any powerlaw density distribution, we find that the sizes (as implied by the visibilities) and the stellar luminosity are incompatible. A reduction to a third of W33A's previously adopted luminosity is required to match the visibilities; such a reduction is consistent with new high resolution 70 micron data from Spitzer's MIPSGAL survey. We obtain best fits for models with shallow dust density distributions of r^(-0.5) and r^(-1.0) and for increased optical depth in the silicate feature produced by decreasing the ISM ratio of graphite to silicates and using optical grain properties by Ossenkopf et al. (1992).Comment: 4 pages, 4 figures. Accepted for ApJ letter

The RMS Survey: The Bolometric Fluxes and Luminosity Distributions of Young Massive Stars

Context: The Red MSX Source (RMS) survey is returning a large sample of massive young stellar objects (MYSOs) and ultra-compact (UC) \HII{} regions using follow-up observations of colour-selected candidates from the MSX point source catalogue. Aims: To obtain the bolometric fluxes and, using kinematic distance information, the luminosities for young RMS sources with far-infrared fluxes. Methods: We use a model spectral energy distribution (SED) fitter to obtain the bolometric flux for our sources, given flux data from our work and the literature. The inputs to the model fitter were optimised by a series of investigations designed to reveal the effect varying these inputs had on the resulting bolometric flux. Kinematic distances derived from molecular line observations were then used to calculate the luminosity of each source. Results: Bolometric fluxes are obtained for 1173 young RMS sources, of which 1069 have uniquely constrained kinematic distances and good SED fits. A comparison of the bolometric fluxes obtained using SED fitting with trapezium rule integration and two component greybody fits was also undertaken, and showed that both produce considerable scatter compared to the method used here. Conclusions: The bolometric flux results allowed us to obtain the luminosity distributions of YSOs and UC\HII{} regions in the RMS sample, which we find to be different. We also find that there are few MYSOs with L $\geq$ 10$^{5}$\lsol{}, despite finding many MYSOs with 10$^{4}$\lsol{} $\geq$ L $\geq$ 10$^{5}$\lsol{}.Comment: 12 pages, 12 figures, 3 tables, accepted to A&A. The full versions of tables 1 and 2 will be available via the CDS upon publicatio

The RMS Survey: 6 cm continuum VLA observations towards candidate massive YSOs in the northern hemisphere

(Abridged) Context: The Red MSX Source (RMS) survey is an ongoing multi-wavelength observational programme designed to return a large, well-selected sample of massive young stellar objects (MYSOs). We have identified $\sim$2000 MYSO candidates located throughout the Galaxy by comparing the colours of MSX and 2MASS point sources to those of known MYSOs. Aims: To identify the populations of UCHII regions and PNe within the sample and examine their Galactic distribution. Method: We have conducted high resolution radio continuum observations at 6 cm towards 659 MYSO candidates in the northern hemisphere (10\degr< l < 250\degr) using the VLA. In addition to these targeted observations we present archival data towards a further 315 RMS sources extracted from a previous VLA survey of the inner Galaxy. Results: We find radio emission towards 272 ($\sim$27% of the observed sample). Using results from other parts of our multi-wavelength survey we separate these RMS-radio associations into two distinct types of objects, classifying 51 as PNe and a further 208 as either compact or UC HII regions. Using this well selected sample of HII regions we estimate their Galactic scale height to be 0.6\degr. Conclusions: Using radio continuum and archival data we have identified 79 PNe and 391 HII regions within the northern RMS catalogue. We estimate the total fraction of contamination by PNe in the RMS sample is of order 10%. The sample of HII regions is probably the best representation to date of the Galactic population of HII regions as a whole.Comment: Accepted for publication in Astronomy and Astrophysics. 15 pages, 9 figures and 5 tables. Full versions of Tables 3, 4 and 5 and Figs. 2, 4 and 7 will only be available via CDS or the RMS website at http:/www.ast.leeds.ac.uk/cgi-bin/RMS/RMS_VLA_IMAGES.cg

The RMS Survey: Far-Infrared Photometry of Young Massive Stars

Context: The Red MSX Source (RMS) survey is a multi-wavelength campaign of follow-up observations of a colour-selected sample of candidate massive young stellar objects (MYSOs) in the galactic plane. This survey is returning the largest well-selected sample of MYSOs to date, while identifying other dust contaminant sources with similar mid-infrared colours including a large number of new ultra-compact (UC)HII regions. Aims:To measure the far-infrared (IR) flux, which lies near the peak of the spectral energy distribution (SED) of MYSOs and UCHII regions, so that, together with distance information, the luminosity of these sources can be obtained. Methods:Less than 50% of RMS sources are associated with IRAS point sources with detections at 60 micron and 100 micron, though the vast majority are visible in Spitzer MIPSGAL or IRAS Galaxy Atlas (IGA) images. However, standard aperture photometry is not appropriate for these data due to crowding of sources and strong spatially variable far-IR background emission in the galactic plane. A new technique using a 2-dimensional fit to the background in an annulus around each source is therefore used to obtain far-IR photometry for young RMS sources. Results:Far-IR fluxes are obtained for a total of 1113 RMS candidates identified as young sources. Of these 734 have flux measurements using IGA 60 micron and 100 micron images and 724 using MIPSGAL 70 micron images, with 345 having measurements in both data sets.Comment: 10 pages, 10 figures, 2 Tables, accepted to A&A. A full version of table 1 is available from the lead author or at the CDS upon publicatio

Water distribution in shocked regions of the NGC1333-IRAS4A protostellar outflow

We present the study of the H2O spatial distribution at two bright shocked regions along IRAS4A, one of the strongest H2O emitters among the Class 0 outflows. We obtained Herschel-PACS maps of the IRAS4A outflow and HIFI observations of two shocked positions. The largest HIFI beam of 38 arcsec at 557 GHz was mapped in several key water lines with different upper energy levels, to reveal possible spatial variations of the line profiles. We detect four H2O lines and CO (16-15) at the two selected positions. In addition, transitions from related outflow and envelope tracers are detected. Different gas components associated with the shock are identified in the H2O emission. In particular, at the head of the red lobe of the outflow, two distinct gas components with different excitation conditions are distinguished in the HIFI emission maps: a compact component, detected in the ground-state water lines, and a more extended one. Assuming that these two components correspond to two different temperature components observed in previous H2O and CO studies, the excitation analysis of the H2O emission suggests that the compact (about 3 arcsec) component is associated with a hot (T~1000 K) gas with densities ~(1-4)x10^5 cm^{-3}, whereas the extended one (10-17 arcsec) traces a warm (T~300-500 K) and dense gas (~(3-5)x10^7 cm^{-3}). Finally, using the CO (16-15) emission observed at R2, we estimate the H2O/H2 abundance of the warm and hot components to be (7-10)x10^{-7} and (3-7)x10^{-5}. Our data allowed us, for the first time, to resolve spatially the two temperature components previously observed with HIFI and PACS. We propose that the compact hot component may be associated with the jet that impacts the surrounding material, whereas the warm, dense, and extended component originates from the compression of the ambient gas by the propagating flow.Comment: 13 pages, 11 figures. Accepted for publication in Astronomy and Astrophysic