Targeted silencing of Jab1/Csn5 in human cells downregulates SCF activity through reduction of F-box protein levels

BACKGROUND: SCF ubiquitin ligases target numerous proteins for ubiquitin dependent proteolysis, including p27 and cyclin E. SCF and other cullin-RING ligases (CRLs) are regulated by the ubiquitin-like protein Nedd8 that covalently modifies the cullin subunit. The removal of Nedd8 is catalyzed by the Jab1/MPN domain metalloenzyme (JAMM) motif within the Csn5 subunit of the Cop9 Signalosome. RESULTS: Here, we conditionally knock down Csn5 expression in HEK293 human cells using a doxycycline-inducible shRNA system. Cullin levels were not altered in CSN-deficient human cells, but the levels of multiple F-box proteins were decreased. Molecular analysis indicates that this decrease was due to increased Cul1- and proteasome-dependent turnover. Diminished F-box levels resulted in reduced SCF activity, as evidenced by accumulation of two substrates of the F-box protein Fbw7, cyclin E and c-myc, in Csn5-depleted cells. CONCLUSION: We propose that deneddylation of Cul1 is required to sustain optimal activity of SCF ubiquitin ligases by repressing 'autoubiquitination' of F-box proteins within SCF complexes, thereby rescuing them from premature degradation

Heat shock factor 1 regulates lifespan as distinct from disease onset in prion disease

Prion diseases are fatal, transmissible, neurodegenerative diseases caused by the misfolding of the prion protein (PrP). At present, the molecular pathways underlying prion-mediated neurotoxicity are largely unknown. We hypothesized that the transcriptional regulator of the stress response, heat shock factor 1 (HSF1), would play an important role in prion disease. Uninoculated HSF1 knockout (KO) mice used in our study do not show signs of neurodegeneration as assessed by survival, motor performance, or histopathology. When inoculated with Rocky Mountain Laboratory (RML) prions HSF1 KO mice had a dramatically shortened lifespan, succumbing to disease ≈20% faster than controls. Surprisingly, both the onset of home-cage behavioral symptoms and pathological alterations occurred at a similar time in HSF1 KO and control mice. The accumulation of proteinase K (PK)-resistant PrP also occurred with similar kinetics and prion infectivity accrued at an equal or slower rate. Thus, HSF1 provides an important protective function that is specifically manifest after the onset of behavioral symptoms of prion disease

Connexin36 knockout mice display increased sensitivity to pentylenetetrazol-induced seizure-like behaviors

Large-scale synchronous firing of neurons during seizures is modulated by electrotonic coupling between neurons via gap junctions. To explore roles for connexin36 (Cx36) gap junctions in seizures, we examined the seizure threshold of connexin36 knockout (Cx36KO) mice using a pentylenetetrazol (PTZ) model

z~2: An Epoch of Disk Assembly

We explore the evolution of the internal gas kinematics of star-forming galaxies from the peak of cosmic star-formation at $z\sim2$ to today. Measurements of galaxy rotation velocity $V_{rot}$, which quantify ordered motions, and gas velocity dispersion $\sigma_g$, which quantify disordered motions, are adopted from the DEEP2 and SIGMA surveys. This sample covers a continuous baseline in redshift from $z=2.5$ to $z=0.1$, spanning 10 Gyrs. At low redshift, nearly all sufficiently massive star-forming galaxies are rotationally supported ($V_{rot}>\sigma_g$). By $z=2$, the percentage of galaxies with rotational support has declined to 50$\%$ at low stellar mass ($10^{9}-10^{10}\,M_{\odot}$) and 70$\%$ at high stellar mass ($10^{10}-10^{11}M_{\odot}$). For $V_{rot}\,>\,3\,\sigma_g$, the percentage drops below 35$\%$ for all masses. From $z\,=\,2$ to now, galaxies exhibit remarkably smooth kinematic evolution on average. All galaxies tend towards rotational support with time, and it is reached earlier in higher mass systems. This is mostly due to an average decline in $\sigma_g$ by a factor of 3 since a redshift of 2, which is independent of mass. Over the same time period, $V_{rot}$ increases by a factor of 1.5 for low mass systems, but does not evolve for high mass systems. These trends in $V_{rot}$ and $\sigma_g$ with time are at a fixed stellar mass and should not be interpreted as evolutionary tracks for galaxy populations. When galaxy populations are linked in time with abundance matching, not only does $\sigma_g$ decline with time as before, but $V_{rot}$ strongly increases with time for all galaxy masses. This enhances the evolution in $V_{rot}/\sigma_g$. These results indicate that $z\,=\,2$ is a period of disk assembly, during which the strong rotational support present in today's massive disk galaxies is only just beginning to emerge.Comment: 12 pages, 8 figures, submitted to Ap

Brainstem Auditory Evoked Potentials' Diagnostic Accuracy for Hearing Loss: Systematic Review and Meta-Analysis

Background: Microvascular decompression (MVD) utilizes brainstem auditory evoked potential (BAEP) intraoperative monitoring to reduce the risk of iatrogenic hearing loss. Studies report varying efficacy and hearing loss rates during MVD with intraoperative monitoring. Objectives: This study aims to perform a comprehensive review and study of diagnostic accuracy of BAEPs during MVD to predict hearing loss in studies published from January 1984 to December 2013. Methods: The PubMed/MEDLINE and World Science databases were searched. Studies performed MVD for trigeminal neuralgia, hemifacial spasm, glossopharyngeal neuralgia or geniculate neuralgia and monitored intraoperative BAEPs to prevent hearing loss. Retrospectively, BAEP parameters were compared with postoperative hearing. The diagnostic accuracy of significant change in BAEPs, which includes loss of response, was tested using summary receiver operative curve and diagnostic odds ratio (DOR). Results: A total of 13 studies were included in the analysis with a total of 2,540 cases. Loss of response pooled sensitivity, specificity, and DOR with 95% confidence interval being 74% (60–84%), 98% (88–100%), and 69.3 (18.2–263%), respectively. The similar significant change results were 88% (77–94%), 63% (40–81%), and 9.1 (3.9–21.6%). Conclusion: Patients with hearing loss after MVD are more likely to have shown loss of BAEP responses intraoperatively. Loss of responses has high specificity in evaluating hearing loss. Patients undergoing MVD should have BAEP monitoring to prevent hearing loss

Attenuated Expression of DFFB is a Hallmark of Oligodendrogliomas with 1p-Allelic Loss

Allelic loss of chromosome 1p is frequently observed in oligodendroglioma. We screened 177 oligodendroglial tumors for 1p deletions and found 6 tumors with localized 1p36 deletions. Several apoptosis regulation genes have been mapped to this region, including Tumor Protein 73 (p73), DNA Fragmentation Factor subunits alpha (DFFA) and beta (DFFB), and Tumor Necrosis Factor Receptor Superfamily Members 9 and 25 (TNFRSF9, TNFRSF25). We compared expression levels of these 5 genes in pairs of 1p-loss and 1p-intact tumors using quantitative reverse-transcriptase PCR (QRTPCR) to test if 1p deletions had an effect on expression. Only the DFFB gene demonstrated decreased expression in all tumor pairs tested. Mutational analysis did not reveal DFFB mutations in 12 tested samples. However, it is possible that DFFB haploinsufficiency from 1p allelic loss is a contributing factor in oligodendroglioma development

Role of Predicted Metalloprotease Motif of Jab1/Csn5 in Cleavage of Nedd8 from Cul1

COP9 signalosome (CSN) cleaves the ubiquitin-like protein Nedd8 from the Cul1 subunit of SCF ubiquitin ligases. The Jab1/MPN domain metalloenzyme (JAMM) motif in the Jab1/Csn5 subunit was found to underlie CSN's Nedd8 isopeptidase activity. JAMM is found in proteins from archaea, bacteria, and eukaryotes, including the Rpn11 subunit of the 26S proteasome. Metal chelators and point mutations within JAMM abolished CSN-dependent cleavage of Nedd8 from Cul1, yet had little effect on CSN complex assembly. Optimal SCF activity in yeast and both viability and proper photoreceptor cell (R cell) development in Drosophila melanogaster required an intact Csn5 JAMM domain. We propose that JAMM isopeptidases play important roles in a variety of physiological pathways

Sedimentary DNA and molecular evidence for early human occupation of the Faroe Islands

The Faroe Islands, a North Atlantic archipelago between Norway and Iceland, were settled by Viking explorers in the mid-9th century CE. However, several indirect lines of evidence suggest earlier occupation of the Faroes by people from the British Isles. Here, we present sedimentary ancient DNA and molecular fecal biomarker evidence from a lake sediment core proximal to a prominent archaeological site in the Faroe Islands to establish the earliest date for the arrival of people in the watershed. Our results reveal an increase in fecal biomarker concentrations and the first appearance of sheep DNA at 500 CE (95% confidence interval 370-610 CE), pre-dating Norse settlements by 300 years. Sedimentary plant DNA indicates an increase in grasses and the disappearance of woody plants, likely due to livestock grazing. This provides unequivocal evidence for human arrival and livestock disturbance in the Faroe Islands centuries before Viking settlement in the 9th century

A Deep Chandra X-ray Spectrum of the Accreting Young Star TW Hydrae

We present X-ray spectral analysis of the accreting young star TW Hydrae from a 489 ks observation using the Chandra High Energy Transmission Grating. The spectrum provides a rich set of diagnostics for electron temperature T_e, electron density N_e, hydrogen column density N_H, relative elemental abundances and velocities and reveals its source in 3 distinct regions of the stellar atmosphere: the stellar corona, the accretion shock, and a very large extended volume of warm postshock plasma. The presence of Mg XII, Si XIII, and Si XIV emission lines in the spectrum requires coronal structures at ~10 MK. Lower temperature lines (e.g., from O VIII, Ne IX, and Mg XI) formed at 2.5 MK appear more consistent with emission from an accretion shock. He-like Ne IX line ratio diagnostics indicate that T_e = 2.50 +/- 0.25 MK and N_e = 3.0 +/- 0.2 x 10^(12) cm^(-3) in the shock. These values agree well with standard magnetic accretion models. However, the Chandra observations significantly diverge from current model predictions for the postshock plasma. This gas is expected to cool radiatively, producing O VII as it flows into an increasingly dense stellar atmosphere. Surprisingly, O VII indicates N_e = 5.7 ^(+4.4}_(-1.2) x 10^(11) cm^(-3), five times lower than N_e in the accretion shock itself, and ~7 times lower than the model prediction. We estimate that the postshock region producing O VII has roughly 300 times larger volume, and 30 times more emitting mass than the shock itself. Apparently, the shocked plasma heats the surrounding stellar atmosphere to soft X-ray emitting temperatures and supplies this material to nearby large magnetic structures -- which may be closed magnetic loops or open magnetic field leading to mass outflow. (Abridged)Comment: 13 pages (emulateapj style), 10 figures, ApJ, in pres