Crystal structure of the catalytic D2 domain of the AAA+ ATPase p97 reveals a putative helical split-washer-type mechanism for substrate unfolding

Several pathologies have been associated with the AAA+ ATPase p97, an enzyme essential to protein homeostasis. Heterozygous polymorphisms in p97 have been shown to cause neurological disease, while elevated proteotoxic stress in tumours has made p97 an attractive cancer chemotherapy target. The cellular processes reliant on p97 are well described. High‐resolution structural models of its catalytic D2 domain, however, have proved elusive, as has the mechanism by which p97 converts the energy from ATP hydrolysis into mechanical force to unfold protein substrates. Here, we describe the high‐resolution structure of the p97 D2 ATPase domain. This crystal system constitutes a valuable tool for p97 inhibitor development and identifies a potentially druggable pocket in the D2 domain. In addition, its P61 symmetry suggests a mechanism for substrate unfolding by p97

Characterization of mixed lymphocyte reaction blocking antibodies (MLR-Bf) in human pregnancy

BACKGROUND: It is known that during normal pregnancy and after immunotherapy blocking antibodies are developed, these antibodies inhibit mixed lymphocyte reaction and are also anti-mitogenic in nature. Mixed lymphocyte reaction blocking antibodies are specific to the husband's lymphocytes. In the present study an attempt has been made to characterize the mixed lymphocyte reaction blocking antibodies in normal pregnancy and in women with recurrent spontaneous abortion after immunotherapy. METHODS: Serum was obtained from women of different gestational windows of pregnancy (Ist, IInd, IIIrd trimesters and post delivery period of normal pregnancy), recurrent spontaneous aborters from pre and post immunization. Healthy (male and females) controls were screened for the presence of mixed lymphocyte reaction blocking antibodies. The standard mixed lymphocyte reaction technique was used to evaluate the inhibitory effect of serum in the mixed lymphocyte reaction. Each serum was tested for cytotoxic antibodies. Immunoglobulin G and its isotypes were isolated according to the standard protocol. RESULTS: In the present study we have observed that there was significant inhibition of proliferation response when immunoglobulin G from different trimesters of pregnancy were added to one way mixed lymphocyte reaction or to phytohemagglutinin activated lymphocyte proliferation assay. Similar pattern was seen when immunoglobulin G isolated from adequately immunized women with recurrent spontaneous abortion was used. It was further confirmed that amongst all the isotypes of immunoglobulin G, only immunoglobulin G-3 was found to be positive for the inhibitory effect. CONCLUSIONS: Present study indicates that mixed lymphocyte reaction blocking antibodies are immunoglobulin G-3 in nature. It is developed during pregnancy and also after immunotherapy in women with recurrent spontaneous abortion who subsequently have the successful pregnancy

Epigenetics of human cutaneous melanoma: setting the stage for new therapeutic strategies

Cutaneous melanoma is a very aggressive neoplasia of melanocytic origin with constantly growing incidence and mortality rates world-wide. Epigenetic modifications (i.e., alterations of genomic DNA methylation patterns, of post-translational modifications of histones, and of microRNA profiles) have been recently identified as playing an important role in melanoma development and progression by affecting key cellular pathways such as cell cycle regulation, cell signalling, differentiation, DNA repair, apoptosis, invasion and immune recognition. In this scenario, pharmacologic inhibition of DNA methyltransferases and/or of histone deacetylases were demonstrated to efficiently restore the expression of aberrantly-silenced genes, thus re-establishing pathway functions. In light of the pleiotropic activities of epigenetic drugs, their use alone or in combination therapies is being strongly suggested, and a particular clinical benefit might be expected from their synergistic activities with chemo-, radio-, and immuno-therapeutic approaches in melanoma patients. On this path, an important improvement would possibly derive from the development of new generation epigenetic drugs characterized by much reduced systemic toxicities, higher bioavailability, and more specific epigenetic effects

The nature of sub-millimetre galaxies II: an ALMA comparison of SMG dust heating mechanisms

We compare the contribution of active galactic nuclei (AGNs) and star formation towards dust heating in sub-mm galaxies (SMGs). We have used ALMA at 0.1-arcsec resolution to image a complete flux-limited sample of seven sub-mm sources previously shown to have spectral energy distributions that were as well-fitted by obscured AGN as star-forming galaxy templates. Indeed, two sub-mm sources were known to be quasars from their absorbed X-ray emission. We find the sub-mm sizes of all SMGs to be small (≈1−2 kpc) and generally ∼3 times smaller than any host detected in the near-infrared (NIR). In all cases, the five SMGs are comparable in sub-mm size to the two known quasars and four z ≈ 6 quasars, also observed with ALMA. We detect no evidence of diffuse spiral arms in this complete sample. We then convert the far-infrared (FIR) luminosities to star formation rate (SFR) surface densities and find that the SMGs occupy the same range as the known quasars in our sample. We conclude that in terms of sub-mm size, extent relative to host and SFR density as well as luminosity and mid-IR (MIR) colour, there is little distinction between the SMGs and sub-mm bright quasars. Finally, we present preliminary evidence that SMGs with higher MIR luminosities and sub-mm loud quasars tend to have dust components that range to hotter temperatures than their less luminous SMG counterparts. In light of these results, we continue to suggest that luminous SMGs may host dust-absorbed quasars that may simultaneously dominate the FIR and hard X-ray backgrounds

KASHz: No evidence for ionised outflows instantaneously suppressing star formation in moderate luminosity AGN at z∼1.4–2.6

As part of our KMOS AGN Survey at High-redshift (KASHz), we present spatially-resolved VLT/KMOS and VLT/SINFONI spectroscopic data and ALMA 870μm continuum imaging of eight z=1.4–2.6 moderate AGN (⁠L 2−10kev L2−10kev = 1042 − 1045 ergs s−1). We map [O iii], Hα and rest-frame FIR emission to search for any spatial anti-correlation between ionised outflows (traced by the [O iii] line) and star formation (SF; traced by Hα and FIR), that has previously been claimed for some high-z AGN and used as evidence for negative and/or positive AGN feedback. Firstly, we conclude that Hα is unreliable to map SF inside our AGN host galaxies based on: (i) SF rates inferred from attenuation-corrected Hα can lie below those inferred from FIR; (ii) the FIR continuum is more compact than the Hα emission by a factor of ∼2 on average; (iii) in half of our sample, we observe significant spatial offsets between the FIR and Hα emission, with an average offset of 1.4 ± 0.6 kpc. Secondly, for the five targets with outflows we find no evidence for a spatial anti-correlation between outflows and SF using either Hα or FIR as a tracer. This holds for our re-analysis of a famous z=1.6 X-ray AGN (‘XID 2028’) where positive and negative feedback has been previously claimed. Based on our results, any impact on SF by ionised outflows must be subtle, either occurring on scales below our resolution, or on long timescales