The melanoma-associated antigen 1 (MAGEA1) protein stimulates the E3 ubiquitin-ligase activity of TRIM31 within a TRIM31-MAGEA1-NSE4 complex

The MAGE (Melanoma-associated antigen) protein family members are structurally related to each other by a MAGEhomology domain comprised of 2 winged helix motifs WH/A and WH/B. This family specifically evolved in placental mammals although single homologs designated NSE3 (non-SMC element) exist in most eukaryotes. NSE3, together with its partner proteins NSE1 and NSE4 form a tight subcomplex of the structural maintenance of chromosomes SMC5–6 complex. Previously, we showed that interactions of the WH/B motif of the MAGE proteins with their NSE4/EID partners are evolutionarily conserved (including the MAGEA1-NSE4 interaction). In contrast, the interaction of the WH/A motif of NSE3 with NSE1 diverged in the MAGE paralogs. We hypothesized that the MAGE paralogs acquired new RING-finger containing partners through their evolution and form MAGE complexes reminiscent of NSE1-NSE3-NSE4 trimers. In this work, we employed the yeast 2-hybrid system to screen a human RING-finger protein library against several MAGE baits. We identified a number of potential MAGE-RING interactions and confirmed several of them (MDM4, PCGF6, RNF166, TRAF6, TRIM8, TRIM31, TRIM41) in co-immunoprecipitation experiments. Among these MAGE-RING pairs, we chose to examine MAGEA1-TRIM31 in detail and showed that both WH/A and WH/B motifs of MAGEA1 bind to the coiled-coil domain of TRIM31 and that MAGEA1 interaction stimulates TRIM31 ubiquitin-ligase activity. In addition, TRIM31 directly binds to NSE4, suggesting the existence of a TRIM31-MAGEA1-NSE4 complex reminiscent of the NSE1-NSE3-NSE4 trimer. These results suggest that MAGEA1 functions as a co-factor of TRIM31 ubiquitin-ligase and that the TRIM31-MAGEA1-NSE4 complex may have evolved from an ancestral NSE1-NSE3-NSE4 complex

Preparing an unsupervised massive analysis of SPHERE high contrast data with the PACO algorithm

We aim at searching for exoplanets on the whole ESO/VLT-SPHERE archive with improved and unsupervised data analysis algorithm that could allow to detect massive giant planets at 5 au. To prepare, test and optimize our approach, we gathered a sample of twenty four solar-type stars observed with SPHERE using angular and spectral differential imaging modes. We use PACO, a new generation algorithm recently developed, that has been shown to outperform classical methods. We also improve the SPHERE pre-reduction pipeline, and optimize the outputs of PACO to enhance the detection performance. We develop custom built spectral prior libraries to optimize the detection capability of the ASDI mode for both IRDIS and IFS. Compared to previous works conducted with more classical algorithms than PACO, the contrast limits we derived are more reliable and significantly better, especially at short angular separations where a gain by a factor ten is obtained between 0.2 and 0.5 arcsec. Under good observing conditions, planets down to 5 MJup, orbiting at 5 au could be detected around stars within 60 parsec. We identified two exoplanet candidates that require follow-up to test for common proper motion. In this work, we demonstrated on a small sample the benefits of PACO in terms of achievable contrast and of control of the confidence levels. Besides, we have developed custom tools to take full benefits of this algorithm and to quantity the total error budget on the estimated astrometry and photometry. This work paves the way towards an end-to-end, homogeneous, and unsupervised massive re-reduction of archival direct imaging surveys in the quest of new exoJupiters.Comment: Accepted for publication in A&

Peering into the Young Planetary System AB Pic. Atmosphere, Orbit, Obliquity & Second Planetary Candidate

We aim to revisit the system AB Pic which has a known companion at the exoplanet/ brown-dwarf boundary. We based this study on a rich set of observations to investigate the companion's orbit and atmosphere. We composed a spectrum of AB Pic b merging archival VLT/SINFONI K-band data, with published spectra at J and H-band (SINFONI) and Lp-band (Magellan-AO), and photometric measurements (HST and Spitzer). We modeled the spectrum with ForMoSA, based on two atmospheric models: ExoREM and BT-SETTL13. We determined the orbital properties of b fitting the astrometric measurements from NaCo (2003 and 2004) and SPHERE (2015). The orbital solutions favor a semi-major axis of $\sim$190au viewed edge-on. With Exo-REM, we derive a T$_{eff}$ of 1700$\pm$50K and surface gravity of 4.5$\pm$0.3dex, consistent with previous works, and we report for the first time a C/O ratio of 0.58$\pm$0.08 ($\sim$solar). The posteriors are sensitive to the wavelength interval and the family of models used. Given the 2.1hr rotation period and our vsin(i) of $\sim$73km/s, we estimate for the first time the true obliquity to be $\sim$45 or $\sim$135deg, indicating a significant misalignment between the planet's spin and orbit orientations. Finally, the existence of a proper motion anomaly between the Hipparcos and Gaia eDR3 compared to our SPHERE detection limits and adapted radial velocity limits indicate the existence of a $\sim$6M$_{Jup}$ inner planet orbiting from 2 to 10au (40-200mas). The possible existence of an inner companion, together with the likely miss-alignment of the spin axis orientation, strongly favor a formation path by gravitational instability or core accretion within a disk closer inside followed by dynamical interactions. Confirmation and characterization of planet c and access to a broader wavelength coverage for planet b will be essential to probe the uncertainties associated with the parameters.Comment: 17 pages, 13 Figures, 6 Tables. Accepted for publication in A&A (31 of October

Interactions between the Nse3 and Nse4 Components of the SMC5-6 Complex Identify Evolutionarily Conserved Interactions between MAGE and EID Families

The SMC5-6 protein complex is involved in the cellular response to DNA damage. It is composed of 6-8 polypeptides, of which Nse1, Nse3 and Nse4 form a tight sub-complex. MAGEG1, the mammalian ortholog of Nse3, is the founding member of the MAGE (melanoma-associated antigen) protein family and Nse4 is related to the EID (E1A-like inhibitor of differentiation) family of transcriptional repressors.Using site-directed mutagenesis, protein-protein interaction analyses and molecular modelling, we have identified a conserved hydrophobic surface on the C-terminal domain of Nse3 that interacts with Nse4 and identified residues in its N-terminal domain that are essential for interaction with Nse1. We show that these interactions are conserved in the human orthologs. Furthermore, interaction of MAGEG1, the mammalian ortholog of Nse3, with NSE4b, one of the mammalian orthologs of Nse4, results in transcriptional co-activation of the nuclear receptor, steroidogenic factor 1 (SF1). In an examination of the evolutionary conservation of the Nse3-Nse4 interactions, we find that several MAGE proteins can interact with at least one of the NSE4/EID proteins.We have found that, despite the evolutionary diversification of the MAGE family, the characteristic hydrophobic surface shared by all MAGE proteins from yeast to humans mediates its binding to NSE4/EID proteins. Our work provides new insights into the interactions, evolution and functions of the enigmatic MAGE proteins

An inner warp discovered in the disk around HD 110058 using VLT/SPHERE and HST/STIS

An edge-on debris disk was detected in 2015 around the young, nearby A0V star HD 110058. The disk showed features resembling those seen in the disk of beta Pictoris that could indicate the presence of a perturbing planetary-mass companion in the system. We investigated new and archival scattered light images of the disk in order to characterise its morphology and spectrum. In particular, we analysed the disk's warp to constrain the properties of possible planetary perturbers. Our work uses data from two VLT/SPHERE observations and archival data from HST/STIS. We measured the morphology of the disk by analysing vertical profiles along the length of the disk to extract the centroid spine position and vertical height. We extracted the surface brightness and reflectance spectrum of the disk. We detect the disk between 20 au (with SPHERE) and 150 au (with STIS), at a position angle of 159.6$^\circ\pm$0.6$^\circ$. Analysis of the spine shows an asymmetry between the two sides of the disk, with a 3.4$^\circ\pm$0.9$^\circ$ warp between ~20 au and 60 au. The disk is marginally vertically resolved in scattered light, with a vertical aspect ratio of 9.3$\pm$0.7% at 45 au. The extracted reflectance spectrum is featureless, flat between 0.95 micron and 1.1 micron, and red from 1.1 micron to 1.65 micron. The outer parts of the disk are also asymmetric with a tilt between the two sides compatible with a disk made of forward-scattering particles and seen not perfectly edge-on, suggesting an inclination of <84$^\circ$. The presence of an undetected planetary-mass companion on an inclined orbit with respect to the disk could explain the warp. The misalignment of the inner parts of the disk with respect to the outer disk suggests a warp that has not yet propagated to the outer parts of the disk, favouring the scenario of an inner perturber as the origin of the warp.Comment: 17 pages, 15 figures, 3 tables; accepted for publication in A&

MAGE-A cancer/testis antigens inhibit MDM2 ubiquitylation function and promote increased levels of MDM4

Melanoma antigen A (MAGE-A) proteins comprise a structurally and biochemically similar sub-family of Cancer/Testis antigens that are expressed in many cancer types and are thought to contribute actively to malignancy. MAGE-A proteins are established regulators of certain cancer-associated transcription factors, including p53, and are activators of several RING finger-dependent ubiquitin E3 ligases. Here, we show that MAGE-A2 associates with MDM2, a ubiquitin E3 ligase that mediates ubiquitylation of more than 20 substrates including mainly p53, MDM2 itself, and MDM4, a potent p53 inhibitor and MDM2 partner that is structurally related to MDM2. We find that MAGE-A2 interacts with MDM2 via the N-terminal p53-binding pocket and the RING finger domain of MDM2 that is required for homo/hetero-dimerization and for E2 ligase interaction. Consistent with these data, we show that MAGE-A2 is a potent inhibitor of the E3 ubiquitin ligase activity of MDM2, yet it does not have any significant effect on p53 turnover mediated by MDM2. Strikingly, however, increased MAGE-A2 expression leads to reduced ubiquitylation and increased levels of MDM4. Similarly, silencing of endogenous MAGE-A expression diminishes MDM4 levels in a manner that can be rescued by the proteasomal inhibitor, bortezomid, and permits increased MDM2/MDM4 association. These data suggest that MAGE-A proteins can: (i) uncouple the ubiquitin ligase and degradation functions of MDM2; (ii) act as potent inhibitors of E3 ligase function; and (iii) regulate the turnover of MDM4. We also find an association between the presence of MAGE-A and increased MDM4 levels in primary breast cancer, suggesting that MAGE-A-dependent control of MDM4 levels has relevance to cancer clinically

An imaged 15Mjup companion within a hierarchical quadruple system

Since 2019, the direct imaging B-star Exoplanet Abundance Study (BEAST) at SPHERE@VLT has been scanning the surroundings of young B-type stars in order to ascertain the ultimate frontiers of giant planet formation. Recently, the $17^{+3}_{-4}$ Myr HIP 81208 was found to host a close-in (~50 au) brown dwarf and a wider (~230 au) late M star around the central 2.6Msun primary. Alongside the continuation of the survey, we are undertaking a complete reanalysis of archival data aimed at improving detection performances so as to uncover additional low-mass companions. We present here a new reduction of the observations of HIP 81208 using PACO ASDI, a recent and powerful algorithm dedicated to processing high-contrast imaging datasets, as well as more classical algorithms and a dedicated PSF-subtraction approach. The combination of different techniques allowed for a reliable extraction of astrometric and photometric parameters. A previously undetected source was recovered at a short separation from the C component of the system. Proper motion analysis provided robust evidence for the gravitational bond of the object to HIP 81208 C. Orbiting C at a distance of ~20 au, this 15Mjup brown dwarf becomes the fourth object of the hierarchical HIP 81208 system. Among the several BEAST stars which are being found to host substellar companions, HIP 81208 stands out as a particularly striking system. As the first stellar binary system with substellar companions around each component ever found by direct imaging, it yields exquisite opportunities for thorough formation and dynamical follow-up studies.Comment: 12 pages, 9 figures, 5 tables. Accepted for publication as a Letter in Astronomy and Astrophysics, section 1. Letters to the Edito

MAGE I Transcription Factors Regulate KAP1 and KRAB Domain Zinc Finger Transcription Factor Mediated Gene Repression

Class I MAGE proteins (MAGE I) are normally expressed only in developing germ cells but are aberrantly expressed in many cancers. They have been shown to promote tumor survival, aggressive growth, and chemoresistance but the underlying mechanisms and MAGE I functions have not been fully elucidated. KRAB domain zinc finger transcription factors (KZNFs) are the largest group of vertebrate transcription factors and regulate neoplastic transformation, tumor suppression, cellular proliferation, and apoptosis. KZNFs bind the KAP1 protein and direct KAP1 to specific DNA sequences where it suppresses gene expression by inducing localized heterochromatin characterized by histone 3 lysine 9 trimethylation (H3me3K9). Discovery that MAGE I proteins also bind to KAP1 prompted us to investigate whether MAGE I can affect KZNF and KAP1 mediated gene regulation. We found that expression of MAGE I proteins, MAGE-A3 or MAGE-C2, relieved repression of a reporter gene by ZNF382, a KZNF with tumor suppressor activity. ChIP of MAGE I (-) HEK293T cells showed KAP1 and H3me3K9 are normally bound to the ID1 gene, a target of ZNF382, but that binding is greatly reduced in the presence of MAGE I proteins. MAGE I expression relieved KAP1 mediated ID1 repression, causing increased expression of ID1 mRNA and ID1 chromatin relaxation characterized by loss of H3me3K9. MAGE I binding to KAP1 also induced ZNF382 poly-ubiquitination and degradation, consistent with loss of ZNF382 leading to decreased KAP1 binding to ID1. In contrast, MAGE I expression caused increased KAP1 binding to Ki67, another KAP1 target gene, with increased H3me3K9 and decreased Ki67 mRNA expression. Since KZNFs are required to direct KAP1 to specific genes, these results show that MAGE I proteins can differentially regulate members of the KZNF family and KAP1 mediated gene repression

Destabilized SMC5/6 complex leads to chromosome breakage syndrome with severe lung disease

The structural maintenance of chromosomes (SMC) family of proteins supports mitotic proliferation, meiosis, and DNA repair to control genomic stability. Impairments in chromosome maintenance are linked to rare chromosome breakage disorders. Here, we have identified a chromosome breakage syndrome associated with severe lung disease in early childhood. Four children from two unrelated kindreds died of severe pulmonary disease during infancy following viral pneumonia with evidence of combined T and B cell immunodeficiency. Whole exome sequencing revealed biallelic missense mutations in the NSMCE3 (also known as NDNL2) gene, which encodes a subunit of the SMC5/6 complex that is essential for DNA damage response and chromosome segregation. The NSMCE3 mutations disrupted interactions within the SMC5/6 complex, leading to destabilization of the complex. Patient cells showed chromosome rearrangements, micronuclei, sensitivity to replication stress and DNA damage, and defective homologous recombination. This work associates missense mutations in NSMCE3 with an autosomal recessive chromosome breakage syndrome that leads to defective T and B cell function and acute respiratory distress syndrome in early childhood