Autoregulation of the human splice factor kinase CLK1 through exon skipping and intron retention

© 2018 Elsevier B.V. Alternative splicing is a key process required for the regulation of gene expression in normal development and physiology. It is regulated by splice factors whose activities are in turn regulated by splice factor kinases and phosphatases. The CDC-like protein kinases are a widespread family of splice factor kinases involved in normal physiology and in several diseases including cancer. In humans they include the CLK1, CLK2, CLK3 and CLK4 genes. The expression of CLK1 is regulated through alternative splicing producing both full-length catalytically active and truncated catalytically inactive isoforms, CLKT1 (arising from exon 4 skipping) and CLKT2 (arising from intron 4 retention). We examined CLK1 alternative splicing in a range of cancer cell lines, and report widespread and highly variable rates of exon 4 skipping and intron 4 retention. We also examined the effect of severe environmental stress including heat shock, osmotic shock, and exposure to the alkaloid drug harmine on CLK1 alternative splicing in DU145 prostate cancer cells. All treatments rapidly reduced exon 4 skipping and intron 4 retention, shifting the balance towards full-length CLK1 expression. We also found that the inhibition of CLK1 with the benzothiazole TG003 reduced exon 4 skipping and intron 4 retention suggesting an autoregulatory mechanism. CLK1 inhibition with TG003 also resulted in modified alternative splicing of five cancer-associated genes

The S. pombe translation initiation factor eIF4G is sumoylated and associates with the SUMO protease Ulp2

SUMO is a small post-translational modifier, that is attached to lysine residues in target proteins. It acts by altering proteinprotein interactions, protein localisation and protein activity. SUMO chains can also act as substrates for ubiquitination, resulting in proteasome-mediated degradation of the target protein. SUMO is removed from target proteins by one of a number of specific proteases. The processes of sumoylation and desumoylation have well documented roles in DNA metabolism and in the maintenance of chromatin structure. To further analyse the role of this modification, we have purified protein complexes containing the S. pombe SUMO protease, Ulp2. These complexes contain proteins required for ribosome biogenesis, RNA stability and protein synthesis. Here we have focussed on two translation initiation factors that we identified as co-purifying with Ulp2, eIF4G and eIF3h. We demonstrate that eIF4G, but not eIF3h, is sumoylated. This modification is increased under conditions that produce cytoplasmic stress granules. Consistent with this we observe partial co-localisation of eIF4G and SUMO in stressed cells. Using HeLa cells, we demonstrate that human eIF4GI is also sumoylated; in vitro studies indicate that human eIF4GI is modified on K1368 and K1588, that are located in the C-terminal eIF4A- and Mnk-binding sites respectively

Impact of opioid therapy on sleep and respiratory patterns in adults with advanced cancer receiving palliative care

Context. In advanced cancer, abnormal sleep patterns may contribute to poor quality of life, but the impact of opioidrelated sleep disorders has not been explored in detail in these patients

Adult Non-Cystic Fibrosis Bronchiectasis Is Characterised by Airway Luminal Th17 Pathway Activation

Copyright © 2015 Chen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.BACKGROUND: Non-cystic fibrosis (CF) bronchiectasis is characterised by chronic airway infection and neutrophilic inflammation, which we hypothesised would be associated with Th17 pathway activation. METHODS: Th17 pathway cytokines were quantified in bronchoalveolar lavage fluid (BALF), and gene expression of IL-17A, IL-1β, IL-8 and IL-23 determined from endobronchial biopsies (EBx) in 41 stable bronchiectasis subjects and 20 healthy controls. Relationships between IL-17A levels and infection status, important clinical measures and subsequent Pseudomonas aeruginosa infection were determined. RESULTS: BALF levels of all Th17 cytokines (median (IQR) pg/mL) were significantly higher in bronchiectasis than control subjects, including IL-17A (1.73 (1.19, 3.23) vs. 0.27 (0.24, 0.35), 95% CI 1.05 to 2.21, p<0.0001) and IL-23 (9.48 (4.79, 15.75) vs. 0.70 (0.43, 1.79), 95% CI 4.68 to 11.21, p<0.0001). However, BALF IL-17A levels were not associated with clinical measures or airway microbiology, nor predictive of subsequent P. aeruginosa infection. Furthermore, gene expression of IL-17A in bronchiectasis EBx did not differ from control. In contrast, gene expression (relative to medians of controls) in bronchiectasis EBx was significantly higher than control for IL1β (4.12 (1.24, 8.05) vs 1 (0.13, 2.95), 95% CI 0.05 to 4.07, p = 0.04) and IL-8 (3.75 (1.64, 11.27) vs 1 (0.54, 3.89), 95% CI 0.32 to 4.87, p = 0.02) and BALF IL-8 and IL-1α levels showed significant relationships with clinical measures and airway microbiology. P. aeruginosa infection was associated with increased levels of IL-8 while Haemophilus influenzae was associated with increased IL-1α. CONCLUSIONS AND CLINICAL RELEVANCE: Established adult non-CF bronchiectasis is characterised by luminal Th17 pathway activation, however this pathway may be relatively less important than activation of non-antigen-specific innate neutrophilic immunity

Studying the Physical Diversity of Late-M Dwarfs with Dynamical Masses

We present a systematic study of the physical properties of late-M dwarfs based on high-quality dynamical mass measurements and near-infrared (NIR) spectra. We use astrometry from Keck NGS and LGS AO imaging to determine orbits for late-M binaries. We find that LP 349-25 (M7.5+M8) is a pair of young brown dwarfs (Mtot = 0.120 Msun) for which Lyon and Tucson evolutionary models jointly predict an age of 140+/-30 Myr. This is consistent with the age of the Pleiades, but at least LP 349-25A defies the empirical Pleiades lithium depletion boundary, implying that the system is in fact older and that evolutionary models underpredict the component luminosities. We find that LHS 1901AB (M6.5+M6.5) is a pair of very low-mass stars (Mtot = 0.194 Msun) with model-derived ages consistent with limits from its lack of activity (> 6 Gyr). Our improved orbit for Gl 569Bab (M8.5+M9) results in a higher mass for this binary (Mtot = 0.140 Msun) compared to previous work (0.125 Msun). We use these masses along with our published results for 2MASS J2206-2047AB (M8+M8) to test four sets of ultracool model atmospheres currently in use. Fitting these models to our NIR integrated-light spectra provides temperature estimates warmer by ~250 K than those derived independently from Dusty evolutionary models given the measured masses and luminosities. We propose that model atmospheres are more likely to be the source of this discrepancy, as it would be difficult to explain a uniform temperature offset over such a wide range of masses, ages, and activity levels in the context of evolutionary models. Our results contrast those of Konopacky et al. as we find an opposite and smaller mass discrepancy from what they report when we adopt their model-testing approach since our Teff estimates from fitting spectra are ~650 K higher than from their fitting of broadband photometry alone.Comment: 53 pages, 12 figures, accepted to Ap

Young Stars and Protostellar Cores near NGC 2023

We investigate the young (proto)stellar population in NGC 2023 and the L 1630 molecular cloud bordering the HII region IC 434, using Spitzer IRAC and MIPS archive data, JCMT SCUBA imaging and spectroscopy as well as targeted BIMA observations of one of the Class 0 protostars, NGC 2023 MM1. We have performed photometry of all IRAC and MIPS images, and used color-color diagrams to identify and classify all young stars seen within a 22'x26' field along the boundary between IC 434 and L 1630. For some stars, which have sufficient optical, IR, and/or sub-millimeter data we have also used the online SED fitting tool for a large 2D archive of axisymmetric radiative transfer models to perform more detailed modeling of the observed SEDs. We identify 5 sub-millimeter cores in our 850 and 450 micron SCUBA images, two of which have embedded class 0 or I protostars. Observations with BIMA are used to refine the position and characteristics of the Class 0 source NGC 2023 MM 1. These observations show that it is embedded in a very cold cloud core, which is strongly enhanced in NH2D. We find that HD 37903 is the most massive member of a cluster with 20 -- 30 PMS stars. We also find smaller groups of PMS stars formed from the Horsehead nebula and another elephant trunk structure to the north of the Horsehead. We refine the spectral classification of HD 37903 to B2 Ve. Our study shows that the expansion of the IC 434 HII region has triggered star formation in some of the dense elephant trunk structures and compressed gas inside the L 1630 molecular cloud. This pre-shock region is seen as a sub-millimeter ridge in which stars have already formed. The cluster associated with NGC 2023 is very young, and has a large fraction of Class I sources.Comment: 21 pages, 11 figures. Accepted for publication in A&A Replaced with higher resolution figure

Orbital Orientations of Exoplanets: HAT-P-4b is Prograde and HAT-P-14b is Retrograde

We present observations of the Rossiter-McLaughlin effect for two exoplanetary systems, revealing the orientations of their orbits relative to the rotation axes of their parent stars. HAT-P-4b is prograde, with a sky-projected spin-orbit angle of lambda = -4.9 +/- 11.9 degrees. In contrast, HAT-P-14b is retrograde, with lambda = 189.1 +/- 5.1 degrees. These results conform with a previously noted pattern among the stellar hosts of close-in giant planets: hotter stars have a wide range of obliquities and cooler stars have low obliquities. This, in turn, suggests that three-body dynamics and tidal dissipation are responsible for the short-period orbits of many exoplanets. In addition, our data revealed a third body in the HAT-P-4 system, which could be a second planet or a companion star.Comment: AJ, in press [8 pages

Influenza epidemiology, vaccine coverage and vaccine effectiveness in sentinel Australian hospitals in 2013: the Influenza Complications Alert Network

The National Influenza Program aims to reduce serious morbidity and mortality from influenza by providing public funding for vaccination to at-risk groups. The Influenza Complications Alert Network (FluCAN) is a sentinel hospital-based surveillance program that operates at 14 sites in all states and territories in Australia. This report summarises the epidemiology of hospitalisations with confirmed influenza, estimates vaccine coverage and influenza vaccine protection against hospitalisation with influenza during the 2013 influenza season. In this observational study, cases were defined as patients admitted to one of the sentinel hospitals, with influenza confirmed by nucleic acid testing. Controls were patients who had acute respiratory illnesses who were test-negative for influenza. Vaccine effectiveness was estimated as 1 minus the odds ratio of vaccination in case patients compared with control patients, after adjusting for known confounders. During the period 5 April to 31 October 2013, 631 patients were admitted with confirmed influenza at the 14 FluCAN sentinel hospitals. Of these, 31% were more than 65 years of age, 9.5% were Indigenous Australians, 4.3% were pregnant and 77% had chronic co-morbidities. Influenza B was detected in 30% of patients. Vaccination coverage was estimated at 81% in patients more than 65 years of age but only 49% in patients aged less than 65 years with chronic comorbidities. Vaccination effectiveness against hospitalisation with influenza was estimated at 50% (95% confidence interval: 33%, 63%,