Position-effect variegation revisited: HUSHing up heterochromatin in human cells.

Much of what we understand about heterochromatin formation in mammals has been extrapolated from forward genetic screens for modifiers of position-effect variegation (PEV) in the fruit fly Drosophila melanogaster. The recent identification of the HUSH (Human Silencing Hub) complex suggests that more recent evolutionary developments contribute to the mechanisms underlying PEV in human cells. Although HUSH-mediated repression also involves heterochromatin spreading through the reading and writing of the repressive H3K9me3 histone modification, clear orthologues of HUSH subunits are not found in Drosophila but are conserved in vertebrates. Here we compare the insights into the mechanisms of PEV derived from genetic screens in the fly, the mouse and in human cells, review what is currently known about the HUSH complex and discuss the implications of HUSH-mediated silencing for viral latency. Future studies will provide mechanistic insight into HUSH complex function and reveal the relationship between HUSH and other epigenetic silencing complexes.This work was funded by a Wellcome Trust Principal Research Fellowship to P.J.L. (084957/Z/08/Z) and a Wellcome Trust PhD studentship to I.A.T. The Cambridge Institute for Medical Research is in receipt of a Wellcome Trust Strategic Award.This is the author accepted manuscript. The final version is available from Wiley via http://dx.doi.org/10.1002/bies.20150018

Diagnosing GORD in Respiratory Medicine

Gastroesophageal reflux disease (GORD) is increasing in prevalence and is highly associated with several lung diseases such as asthma and COPD. Current diagnostic methods are imperfect, being insensitive, non-specific, expensive, or invasive. An accurate diagnosis of GORD can aid effective treatment and have significant clinical impact. Novel methods such as exhaled breath condensate analysis and electronic nose technology have the potential to improve the accuracy of diagnosing GORD

ATF7IP-Mediated Stabilization of the Histone Methyltransferase SETDB1 Is Essential for Heterochromatin Formation by the HUSH Complex

The histone methyltransferase SETDB1 plays a central role in repressive chromatin processes, but the functional requirement for its binding partner ATF7IP has remained enigmatic. Here, we show that ATF7IP is essential for SETDB1 stability: nuclear SETDB1 protein is degraded by the proteasome upon ablation of ATF7IP. As a result, ATF7IP is critical for repression that requires H3K9 trimethylation by SETDB1, including transgene silencing by the HUSH complex. Furthermore, we show that loss of ATF7IP phenocopies loss of SETDB1 in genome-wide assays. ATF7IP and SETDB1 knockout cells exhibit near-identical defects in the global deposition of H3K9me3, which results in similar dysregulation of the transcriptome. Overall, these data identify a critical functional role for ATF7IP in heterochromatin formation by regulating SETDB1 abundance in the nucleus.This work was supported by the Wellcome Trust through a Principal Research Fellowship to P.J.L. (101835/Z/13/Z) and a Ph.D. studentship to I.A.T. The CIMR is in receipt of a Wellcome Trust strategic award.This is the final version of the article. It first appeared from Elsevier (Cell Press) via https://doi.org/10.1016/j.celrep.2016.09.05

A Genetic Screen Identifies a Critical Role for the WDR81-WDR91 Complex in the Trafficking and Degradation of Tetherin.

Tetherin (BST2/CD317) is a viral restriction factor that anchors enveloped viruses to host cells and limits viral spread. The HIV-1 Vpu accessory protein counteracts tetherin by decreasing its cell surface expression and targeting it for ubiquitin-dependent endolysosomal degradation. Although the Vpu-mediated downregulation of tetherin has been extensively studied, the molecular details are not completely elucidated. We therefore used a forward genetic screen in human haploid KBM7 cells to identify novel genes required for tetherin trafficking. Our screen identified WDR81 as a novel gene required for tetherin trafficking and degradation in both the presence and absence of Vpu. WDR81 is a BEACH-domain containing protein that is also required for the degradation of EGF-stimulated epidermal growth factor receptor (EGFR) and functions in a complex with the WDR91 protein. In the absence of WDR81 the endolysosomal compartment appears swollen, with enlarged early and late endosomes and reduced delivery of endocytosed dextran to cathepsin-active lysosomes. Our data suggest a role for the WDR81-WDR91 complex in the fusion of endolysosomal compartments and the absence of WDR81 leads to impaired receptor trafficking and degradation.This work was supported by the Wellcome Trust, through a Principal Research Fellowship to PJL (084957/Z/08/Z) and Ph.D studentship to RR (079895/Z/06/Z), by MRC research grant MR/M010007/1 to JPL and by a BBSRC industrial CASE studentship with GSK Research and Development Ltd to LJD. The CIMR is in receipt of a Wellcome Trust strategic award 100140.This is the final version of the article. It first appeared from Wiley via https://doi.org/10.1111/tra.1240

ATF7IP-Mediated Stabilization of the Histone Methyltransferase SETDB1 Is Essential for Heterochromatin Formation by the HUSH Complex.

The histone methyltransferase SETDB1 plays a central role in repressive chromatin processes, but the functional requirement for its binding partner ATF7IP has remained enigmatic. Here, we show that ATF7IP is essential for SETDB1 stability: nuclear SETDB1 protein is degraded by the proteasome upon ablation of ATF7IP. As a result, ATF7IP is critical for repression that requires H3K9 trimethylation by SETDB1, including transgene silencing by the HUSH complex. Furthermore, we show that loss of ATF7IP phenocopies loss of SETDB1 in genome-wide assays. ATF7IP and SETDB1 knockout cells exhibit near-identical defects in the global deposition of H3K9me3, which results in similar dysregulation of the transcriptome. Overall, these data identify a critical functional role for ATF7IP in heterochromatin formation by regulating SETDB1 abundance in the nucleus.This work was supported by the Wellcome Trust through a Principal Research Fellowship to P.J.L. (101835/Z/13/Z) and a Ph.D. studentship to I.A.T. The CIMR is in receipt of a Wellcome Trust strategic award.This is the final version of the article. It first appeared from Elsevier (Cell Press) via https://doi.org/10.1016/j.celrep.2016.09.05

On the timing of retreat of the Loch Lomond (‘Younger Dryas’) Readvance icefield in the SW Scottish Highlands and its wider significance

It has long been assumed that the last glacier expansion in the Scottish Highlands, the Loch Lomond Readvance (LLR), resulted from a cold reversal that was broadly coeval with the ‘Younger Dryas’ episode. This view has recently been challenged, with the suggestion that glacier ice had disappeared from Rannoch Moor, one of the main ice accumulation centres in the SW Scottish Highlands, by as early as 12.5 ka, i.e. within the first half of the ‘Younger Dryas’. Here we present new radiocarbon, tephrostratigraphical and pollen-stratigraphical evidence from one of the key sites on Rannoch Moor, the results of an experiment designed to test this hypothesis. Our results not only contradict that concept, but are fully compatible with other evidence from the SW Scottish Highlands that suggests that the LLR glaciers in this area continued to expand until towards the end of the ‘Younger Dryas’ period, and may have persisted in some places after the onset of the Holocene. We consider the possible reasons for this marked divergence in chronology, a matter that is crucial to resolve because the precise timing of the demise of the LLR glaciers has important palaeoclimatic and other implications. In the wider context, we also draw attention to problems with the general use of the term ‘Younger Dryas’ and why we regard the Greenland stratotype unit and term ‘Greenland Stadial 1’ (GS-1) a more secure stratigraphic comparator

Using foresight futures and systems thinking to evaluate digitally enhanced advanced service concepts for a rolling stock company (ROSCO)

Purpose: This paper reports on a study in conjunction with a UK-based rolling stock leasing company (ROSCO). The aim was to generate and evaluate future operational concepts for digitally enhanced advanced services from the point of view of a ROSCO – one of many stakeholders (or actors) within a future wider mobility ecosystem.Design/Methodology/Approach: The research design followed the Generic Foresight Process Framework (Voros 2003). Desk-based research and horizon scanning analysis revealed technologies, mobility and transport trends, and other predictions towards 2060. A workshop was developed and participants were presented with a series of future scenarios and design fictions for end-to-end intermodal mobility and passenger carbon quotas. A future Mobility Servitization Systems Architecture was developed.Findings: Five future megatrends were identified; Decarbonisation, changing traveller needs, digitisation, mobility ecosystems and new business models in digital ecosystems. The ‘what-if’ activities revealed insights into alternate futures; revealing system of systems (SoS) actors, the role of a ROSCO, integrations, assumptions and operational constraints.Originality/Value: This research contributes to engineering and design methods for digitally enhanced advanced services, particularly for corporate strategic foresight in a dominant design industry. The Mobility Servitization Systems Architecture was seen to be a powerful model for ecosystem understanding.</div