Investigation of PARP-1, PARP-2, and PARG interactomes by affinity-purification mass spectrometry

<p>Abstract</p> <p>Background</p> <p>Poly(ADP-ribose) polymerases (PARPs) catalyze the formation of poly(ADP-ribose) (pADPr), a post-translational modification involved in several important biological processes, namely surveillance of genome integrity, cell cycle progression, initiation of the DNA damage response, apoptosis, and regulation of transcription. Poly(ADP-ribose) glycohydrolase (PARG), on the other hand, catabolizes pADPr and thereby accounts for the transient nature of poly(ADP-ribosyl)ation. Our investigation of the interactomes of PARP-1, PARP-2, and PARG by affinity-purification mass spectrometry (AP-MS) aimed, on the one hand, to confirm current knowledge on these interactomes and, on the other hand, to discover new protein partners which could offer insights into PARPs and PARG functions.</p> <p>Results</p> <p>PARP-1, PARP-2, and PARG were immunoprecipitated from human cells, and pulled-down proteins were separated by gel electrophoresis prior to in-gel trypsin digestion. Peptides were identified by tandem mass spectrometry. Our AP-MS experiments resulted in the identifications of 179 interactions, 139 of which are novel interactions. Gene Ontology analysis of the identified protein interactors points to five biological processes in which PARP-1, PARP-2 and PARG may be involved: RNA metabolism for PARP-1, PARP-2 and PARG; DNA repair and apoptosis for PARP-1 and PARP-2; and glycolysis and cell cycle for PARP-1.</p> <p>Conclusions</p> <p>This study reveals several novel protein partners for PARP-1, PARP-2 and PARG. It provides a global view of the interactomes of these proteins as well as a roadmap to establish the systems biology of poly(ADP-ribose) metabolism.</p

X-ray emission from the giant magnetosphere of the magnetic O-type star NGC 1624-2

We observed NGC 1624-2, the O-type star with the largest known magnetic field Bp~20 kG), in X-rays with the ACIS-S camera onboard the Chandra X-ray Observatory. Our two observations were obtained at the minimum and maximum of the periodic Halpha emission cycle, corresponding to the rotational phases where the magnetic field is the closest to equator-on and pole-on, respectively. With these observations, we aim to characterise the star's magnetosphere via the X-ray emission produced by magnetically confined wind shocks. Our main findings are: (i) The observed spectrum of NGC 1624-2 is hard, similar to the magnetic O-type star Theta 1 Ori C, with only a few photons detected below 0.8 keV. The emergent X-ray flux is 30% lower at the Halpha minimum phase. (ii) Our modelling indicated that this seemingly hard spectrum is in fact a consequence of relatively soft intrinsic emission, similar to other magnetic Of?p stars, combined with a large amount of local absorption (~1-3 x 10^22 cm^-2). This combination is necessary to reproduce both the prominent Mg and Si spectral features, and the lack of flux at low energies. NGC 1624-2 is intrinsically luminous in X-rays (log LX emission ~ 33.4) but 70-95% of the X-ray emission produced by magnetically confined wind shocks is absorbed before it escapes the magnetosphere (log LX ISM corrected ~ 32.5). (iii) The high X-ray luminosity, its variation with stellar rotation, and its large attenuation are all consistent with a large dynamical magnetosphere with magnetically confined wind shocks.Comment: Accepted in MNRAS 13 pages, 10 figures, 4 table

Impact of oral melatonin on the electroretinogram cone response

<p>Abstract</p> <p>Background</p> <p>In the eye, melatonin plays a role in promoting light sensitivity at night and modulating many aspects of circadian retinal physiology. It is also an inhibitor of retinal dopamine, which is a promoter of day vision through the cone system. Consequently, it is possible that oral melatonin (an inhibitor of retinal dopamine) taken to alleviate circadian disorders may affect cone functioning. Our aim was to assess the impact of melatonin on the cone response of the human retina using electroretinography (ERG).</p> <p>Methods</p> <p>Twelve healthy participants aged between 18 to 52 years old were submitted to a placebo-controlled, double-blind, crossover, and counterbalanced-order design. The subjects were tested on 2 sessions beginning first with a baseline ERG, followed by the administration of the placebo or melatonin condition and then, 30 min later, a second ERG to test the effect.</p> <p>Results</p> <p>Following oral melatonin administration, a significant decrease of about 8% of the cone maximal response was observed (mean 6.9 μV ± SEM 2.0; P = 0.0065) along with a prolonged b-wave implicit time of 0.4 ms ± 0.1, 50 minutes after ingestion.</p> <p>Conclusion</p> <p>Oral melatonin appears to reach the eye through the circulation. When it is administered at a time of day when it is not usually present, melatonin appears to reduce input to retinal cones. We believe that the impact of melatonin on retinal function should be taken into consideration when used without supervision in chronic self-medication for sleep or circadian disorder treatment.</p

Comparative proteome analysis of human epithelial ovarian cancer

<p>Abstract</p> <p>Background</p> <p>Epithelial ovarian cancer is a devastating disease associated with low survival prognosis mainly because of the lack of early detection markers and the asymptomatic nature of the cancer until late stage. Using two complementary proteomics approaches, a differential protein expression profile was carried out between low and highly transformed epithelial ovarian cancer cell lines which realistically mimic the phenotypic changes observed during evolution of a tumour metastasis. This investigation was aimed at a better understanding of the molecular mechanisms underlying differentiation, proliferation and neoplastic progression of ovarian cancer.</p> <p>Results</p> <p>The quantitative profiling of epithelial ovarian cancer model cell lines TOV-81D and TOV-112D generated using iTRAQ analysis and two-dimensional electrophoresis coupled to liquid chromatography tandem mass spectrometry revealed some proteins with altered expression levels. Several of these proteins have been the object of interest in cancer research but others were unrecognized as differentially expressed in a context of ovarian cancer. Among these, series of proteins involved in transcriptional activity, cellular metabolism, cell adhesion or motility and cytoskeleton organization were identified, suggesting their possible role in the emergence of oncogenic pathways leading to aggressive cellular behavior.</p> <p>Conclusion</p> <p>The differential protein expression profile generated by the two proteomics approaches combined to complementary characterizations studies will open the way to more exhaustive and systematic representation of the disease and will provide valuable information that may be helpful to uncover the molecular mechanisms related to epithelial ovarian cancer.</p

Testing magnetically confined wind shock models for Beta Cep using XMM-Newton and Chandra phase-resolved X-ray observations

(abridged) We have performed a set of phase-resolved X-ray observations of the magnetic B star Beta Cep, for which theoretical models predict the presence of a confined wind emitting X-rays from stationary shocks. We obtained four observations spaced in rotational phase with XMM-Newton and with Chandra. A detailed analysis of the data was performed to derive both photometric and spectral parameters from the EPIC data, searching for rotational modulation, and to derive the location of the X-ray plasma from the line ratios in the He-like triplets of N, O and Ne from the RGS data. The LETG data were used to constrain the presence of bulk motions in the plasma. The strong rotational modulation predicted by the early, static magnetically confined wind model for the X-ray emission is not observed in Beta Cep. The small modulation present goes in the opposite direction, pointing to the absence of any optically thick disk of neutral material, and showing a modulation consistent with the later, dynamic models of magnetically confined wind models in B stars. The lack of observed bulk motion points to the plasma being confined by a magnetic field, but the low plasma temperature and lack of any flaring show that the plasma is not heated by magnetic reconnection. Therefore, the observations point to X-ray emission from shocks in a magnetically confined wind, with no evidence of an optically thick, dense disk at the magnetic equatorComment: Accepted for publication in Astronomy & Astrophysic

The potential of major ion chemistry to assess groundwater vulnerability of a regional aquifer in southern Quebec (Canada)

Groundwater vulnerability mapping provides useful but limited information for developing protection plans of the resource. Classical vulnerability ranking methods often do not take into account complex hydrostratigraphy and never consider groundwater flow dynamics. The objective of this work was to test the potential of major ion chemistry to assess regional-scale intrinsic groundwater vulnerability. Because it reflects water–sediment and water–rock interactions, the new vulnerability index reflects both infiltration processes and groundwater hydrodynamics. The method was applied on a regional fractured bedrock aquifer located in the Becancour region of southern Quebec (Canada). In this region, hydrogeochemistry shows that freshly recharged groundwater evolves from (Ca, Mg)–HCO3 and Ca–SO4 to Na–HCO3 type with gradually increasing confinement conditions in the fractured aquifer and tends to Na–Cl type locally by mixing with trapped marine pore-water. The new method identified recharge areas as those of the highest vulnerability and gradually decreasing vulnerability as confinement of the aquifer increased. It also highlights local discontinuities in confinement that differ from the regional pattern. Results showed a good correlation between groundwater vulnerability estimated with the new method and nitrate occurrence in groundwater. Eighty-two per cent of all samples presenting detectable nitrate concentrations were characterized by a Hydrogeochemical Vulnerability Index greater than 9 (maximum is 10). The ability of the new vulnerability method to identify areas vulnerable to detectable nitrate concentrations was much higher than that deriving from the DRASTIC method. This work confirms that major ions chemistry contains significant information about groundwater vulnerability and could be used to improve groundwater resource management

Global X-ray properties of the O and B stars in Carina

The key empirical property of the X-ray emission from O stars is a strong correlation between the bolometric and X-ray luminosities. In the framework of the Chandra Carina Complex Project, 129 O and B stars have been detected as X-ray sources; 78 of those, all with spectral type earlier than B3, have enough counts for at least a rough X-ray spectral characterization. This leads to an estimate of the Lx/Lbol ratio for an exceptional number of 60 O stars belonging to the same region and triples the number of Carina massive stars studied spectroscopically in X-rays. The derived log(Lx/Lbol) is -7.26 for single objects, with a dispersion of only 0.21dex. Using the properties of hot massive stars listed in the literature, we compare the X-ray luminosities of different types of objects. In the case of O stars, the Lx/Lbol ratios are similar for bright and faint objects, as well as for stars of different luminosity classes or spectral types. Binaries appear only slightly harder and slightly more luminous in X-rays than single objects; the differences are not formally significant (at the 1% level), except for the Lx/Lbol ratio in the medium (1.0--2.5keV) energy band. Weak-wind objects have similar X-ray luminosities but they display slightly softer spectra compared to "normal" O stars with the same bolometric luminosity. Discarding three overluminous objects, we find a very shallow trend of harder emission in brighter objects. The properties of the few B stars bright enough to yield some spectral information appear to be different overall (constant X-ray luminosities, harder spectra), hinting that another mechanism for producing X-rays, besides wind shocks, might be at work. However, it must be stressed that the earliest and X-ray brightest amongst these few detected objects are similar to the latest O stars, suggesting a possibly smooth transition between the two processes.Comment: 30 pages, 9 figures. Accepted for the ApJS Special Issue on the Chandra Carina Complex Project (CCCP), scheduled for publication in May 2011. All 16 CCCP Special Issue papers are available at http://cochise.astro.psu.edu/Carina_public/special_issue.html through 2011 at leas

Discovery Of A Magnetic Field In The Rapidly Rotating O-Type Secondary Of The Colliding-Wind Binary HD 47129 (Plaskett\u27s Star)

We report the detection of a strong, organized magnetic field in the secondary component of the massive O8III/I+O7.5V/III double-lined spectroscopic binary system HD 47129 (Plaskett\u27s star) in the context of the Magnetism in Massive Stars survey. Eight independent Stokes V observations were acquired using the Echelle SpectroPolarimetric Device for the Observations of Stars (ESPaDOnS) spectropolarimeter at the Canada-France-Hawaii Telescope and the Narval spectropolarimeter at the Telescope Bernard Lyot. Using least-squares deconvolution we obtain definite detections of signal in Stokes V in three observations. No significant signal is detected in the diagnostic null (N) spectra. The Zeeman signatures are broad and track the radial velocity of the secondary component; we therefore conclude that the rapidly rotating secondary component is the magnetized star. Correcting the polarized spectra for the line and continuum of the (sharp-lined) primary, we measured the longitudinal magnetic field from each observation. The longitudinal field of the secondary is variable and exhibits extreme values of -810 +/- 150 and +680 +/- 190 G, implying a minimum surface dipole polar strength of 2850 +/- 500 G. In contrast, we derive an upper limit (3 sigma) to the primary\u27s surface magnetic field of 230 G. The combination of a strong magnetic field and rapid rotation leads us to conclude that the secondary hosts a centrifugal magnetosphere fed through a magnetically confined wind. We revisit the properties of the optical line profiles and X-ray emission - previously interpreted as a consequence of colliding stellar winds - in this context. We conclude that HD 47129 represents a heretofore unique stellar system - a close, massive binary with a rapidly rotating, magnetized component - that will be a rich target for further study