Engrailed cooperates directly with Extradenticle and Homothorax on a distinct class of homeodomain binding sites to repress sloppy paired.

Even skipped (Eve) and Engrailed (En) are homeodomain-containing transcriptional repressors with similar DNA binding specificities that are sequentially expressed in Drosophila embryos. The sloppy-paired (slp) locus is a target of repression by both Eve and En. At blastoderm, Eve is expressed in 7 stripes that restrict the posterior border of slp stripes, allowing engrailed (en) gene expression to be initiated in odd-numbered parasegments. En, in turn, prevents expansion of slp stripes after Eve is turned off. Prior studies showed that the two tandem slp transcription units are regulated by cis-regulatory modules (CRMs) with activities that overlap in space and time. An array of CRMs that generate 7 stripes at blastoderm, and later 14 stripes, surround slp1 (Fujioka and Jaynes, 2012). Surprisingly given their similarity in DNA binding specificity and function, responsiveness to ectopic Eve and En indicates that most of their direct target sites are either in distinct CRMs, or in different parts of coregulated CRMs. We localized cooperative binding sites for En, with the homeodomain-containing Hox cofactors Extradenticle (Exd) and Homothorax (Hth), within two CRMs that drive similar expression patterns. Functional analysis revealed two distinct, redundant sites within one CRM. The other CRM contains a single cooperative site that is both necessary and sufficient for repression in the en domain. Correlating in vivo and in vitro analysis suggests that cooperativity with Exd and Hth is a key ingredient in the mechanism of En-dependent repression, and that apparent affinity in vitro is an unreliable predictor of in vivo function

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

Impacts of a Changing Climate and Land Use on Reindeer Pastoralism: Indigenous Knowledge and Remote Sensing

The Arctic is home to many indigenous peoples, including those who depend on reindeer herding for their livelihood, in one of the harshest environments in the world. For the largely nomadic peoples, reindeer not only form a substantial part of the Arctic food base and economy, but they are also culturally important, shaping their way of life, mythologies, festivals and ceremonies. Reindeer pastoralism or husbandry has been practiced by numerous peoples all across Eurasia for thousands of years and involves moving herds of reindeer, which are very docile animals, from pasture to pasture depending on the season. Thus, herders must adapt on a daily basis to find optimal conditions for their herds according to the constantly changing conditions. Climate change and variability plus rapid development are increasingly creating major changes in the physical environment, ecology, and cultures of these indigenous reindeer herder communities in the North, and climate changes are occurring significantly faster in the Arctic than the rest of the globe, with correspondingly dramatic impacts (Oskal, 2008). In response to these changes, Eurasian reindeer herders have created the EALAT project, a comprehensive new initiative to study these impacts and to develop local adaptation strategies based upon their traditional knowledge of the land and its uses - in targeted partnership with the science and remote sensing community - involving extensive collaborations and coproduction of knowledge to minimize the impacts of the various changes. This chapter provides background on climate and development challenges to reindeer husbandry across the Arctic and an overview of the EALAT initiative, with an emphasis on indigenous knowledge, remote sensing, Geographic Information Systems (GIS), and other scientific data to 'co-produce' datasets for use by herders for improved decision-making and herd management. It also provides a description of the EALAT monitoring data integration and sharing system and portal being developed for reindeer pastoralism. In addition, the chapter provides some preliminary results from the EALAT Project, including some early remote sensing research results

Eurasian Reindeer Pastoralism in a Changing Climate: Indigenous Knowledge and NASA Remote Sensing

It is intended that Reindeer Mapper/EALAT will be able to provide reindeer herders with an efficient tool for managing the real-time movements and migrations of their herds through enabling improved efficiency in linking different members of the herder settlements or communities and providing real-time local, satellite or other data (e.g., ice melt in lakes and rivers, weather events), thus enabling real time adjustments to herd movements to avoid problems such as changing weather/climate conditions, freeze-thaw "lock-out" problems, or take advantage of availability of better pasturelands along migration routes. The system is being designed to incorporate local data to allow users to bring their own data into the system for analysis in addition to the data provided by the system itself. With the local information of the population, up to date environmental data and habitat characteristics, the system could generate maps depicting important features of interest for reindeer managers. One of the products derived from the planned Reindeer Mapper system will be a web-based graphic display that allows analysts to quickly pinpoint areas of interest such as those with large concentrations of reindeer and provide surrounding environmental information. The system could be automatically updated with near-real-time information such as hourly precipitation and snowfall rate and accumulation, daily surface and air temperatures, and vegetation cover conditions. The system could bring attention to the proximity of human and animal populations as part of the need for control response. A local GIS will bring these many layers together with several supporting models, showing only a straightforward graphic of the real-time situation in the field. Because the system proposed will be operating in the Internet environment, it should be virtually accessible from any network computers and wireless remote access from the field. The International Center for Reindeer Husbandry in Kautokeino, Norway, is providing regional and international coordination of and access to data sets and expertise, and will act as overall clearinghouse for EALAT information

Extradenticle and Homothorax Control Adult Muscle Fiber Identity in Drosophila

SummaryHere we identify a key role for the homeodomain proteins Extradenticle (Exd) and Homothorax (Hth) in the specification of muscle fiber fate in Drosophila. exd and hth are expressed in the fibrillar indirect flight muscles but not in tubular jump muscles, and manipulating exd or hth expression converts one muscle type into the other. In the flight muscles, exd and hth are genetically upstream of another muscle identity gene, salm, and are direct transcriptional regulators of the signature flight muscle structural gene, Actin88F. Exd and Hth also impact muscle identity in other somatic muscles of the body by cooperating with Hox factors. Because mammalian orthologs of exd and hth also contribute to muscle gene regulation, our studies suggest that an evolutionarily conserved genetic pathway determines muscle fiber differentiation

Insights into Hox Protein Function from a Large Scale Combinatorial Analysis of Protein Domains

Protein function is encoded within protein sequence and protein domains. However, how protein domains cooperate within a protein to modulate overall activity and how this impacts functional diversification at the molecular and organism levels remains largely unaddressed. Focusing on three domains of the central class Drosophila Hox transcription factor AbdominalA (AbdA), we used combinatorial domain mutations and most known AbdA developmental functions as biological readouts to investigate how protein domains collectively shape protein activity. The results uncover redundancy, interactivity, and multifunctionality of protein domains as salient features underlying overall AbdA protein activity, providing means to apprehend functional diversity and accounting for the robustness of Hox-controlled developmental programs. Importantly, the results highlight context-dependency in protein domain usage and interaction, allowing major modifications in domains to be tolerated without general functional loss. The non-pleoitropic effect of domain mutation suggests that protein modification may contribute more broadly to molecular changes underlying morphological diversification during evolution, so far thought to rely largely on modification in gene cis-regulatory sequences

Leadership Competencies for Digital Transformation : Evidence from Multiple Cases

Digital transformation (DT) is disrupting industrial organizations, which require significant changes in their properties to remain competitive. This calls for strong leadership to drive this transformation. However, what leadership competencies are required to lead DT impactfully is unclear. Therefore, we seek to identify the key leadership competencies by employing a qualitative, grounded theory approach. By conducting interviews of ten DT experts from two hardcore industrial organizations, we highlight five key leadership competencies that industrial organizations need to develop in their leaders: digital vision, digital knowledge, failing fast, empowerment, and managing diverse teams. The results of this study will help industrial organizations to strategically prepare their leadership for the requirements of DT.©2020 Springer. This is a post-peer-review, pre-copyedit version of an article published in Advances in Human Factors, Business Management and Leadership: Proceedings of the AHFE 2020 Virtual Conferences on Human Factors, Business Management and Society, and Human Factors in Management and Leadership, July 16-20, 2020, USA. The final authenticated version is available online at: http://dx.doi.org/0.1007/978-3-030-50791-6_11.fi=vertaisarvioitu|en=peerReviewed

Regulation of Ubx Expression by Epigenetic Enhancer Silencing in Response to Ubx Levels and Genetic Variation

For gene products that must be present in cells at defined concentrations, expression levels must be tightly controlled to ensure robustness against environmental, genetic, and developmental noise. By studying the regulation of the concentration-sensitive Drosophila melanogaster Hox gene Ultrabithorax (Ubx), we found that Ubx enhancer activities respond to both increases in Ubx levels and genetic background. Large, transient increases in Ubx levels are capable of silencing all enhancer input into Ubx transcription, resulting in the complete silencing of this gene. Small increases in Ubx levels, brought about by duplications of the Ubx locus, cause sporadic silencing of subsets of Ubx enhancers. Ubx enhancer silencing can also be induced by outcrossing laboratory stocks to D. melanogaster strains established from wild flies from around the world. These results suggest that enhancer activities are not rigidly determined, but instead are sensitive to genetic background. Together, these findings suggest that enhancer silencing may be used to maintain gene product levels within the correct range in response to natural genetic variation

Contribution of Distinct Homeodomain DNA Binding Specificities to Drosophila Embryonic Mesodermal Cell-Specific Gene Expression Programs

Homeodomain (HD) proteins are a large family of evolutionarily conserved transcription factors (TFs) having diverse developmental functions, often acting within the same cell types, yet many members of this family paradoxically recognize similar DNA sequences. Thus, with multiple family members having the potential to recognize the same DNA sequences in cis-regulatory elements, it is difficult to ascertain the role of an individual HD or a subclass of HDs in mediating a particular developmental function. To investigate this problem, we focused our studies on the Drosophila embryonic mesoderm where HD TFs are required to establish not only segmental identities (such as the Hox TFs), but also tissue and cell fate specification and differentiation (such as the NK-2 HDs, Six HDs and identity HDs (I-HDs)). Here we utilized the complete spectrum of DNA binding specificities determined by protein binding microarrays (PBMs) for a diverse collection of HDs to modify the nucleotide sequences of numerous mesodermal enhancers to be recognized by either no or a single subclass of HDs, and subsequently assayed the consequences of these changes on enhancer function in transgenic reporter assays. These studies show that individual mesodermal enhancers receive separate transcriptional input from both I–HD and Hox subclasses of HDs. In addition, we demonstrate that enhancers regulating upstream components of the mesodermal regulatory network are targeted by the Six class of HDs. Finally, we establish the necessity of NK-2 HD binding sequences to activate gene expression in multiple mesodermal tissues, supporting a potential role for the NK-2 HD TF Tinman (Tin) as a pioneer factor that cooperates with other factors to regulate cell-specific gene expression programs. Collectively, these results underscore the critical role played by HDs of multiple subclasses in inducing the unique genetic programs of individual mesodermal cells, and in coordinating the gene regulatory networks directing mesoderm development.National Institutes of Health (U.S.) (Grant R01 HG005287

Therapeutic jurisprudence and procedural justice in Scottish drug courts

Scotland, like other Western jurisdictions, has recently witnessed the development of problem-solving courts aimed at responding more effectively to issues that underlie certain types of offending behaviour. The first to be established were two pilot Drug Courts which drew upon experience of Scottish Drug Treatment and Testing Orders. In common with Drug Courts elsewhere, the Scottish pilots combined treatment, drug testing, supervision and judicial oversight. This article focuses upon the role of judicial involvement in the ongoing review of Drug Court participants’ progress, drawing upon court observation and interviews with offenders and Drug Court professionals. Drug Court dialogues were typically encouraging on the part of sheriffs, aimed at recognising and reinforcing the progress made by participants and motivating then to maintain and build upon their achievements to date, while participants were generally responsive to the positive feedback they received from the sheriffs as their orders progressed. Interactions within the Scottish Drug Courts reflect key features of procedural justice (Tyler, 1990), including ethicality, efforts to be fair and representation. By contributing to enhanced perceptions of procedural justice, Drug Court dialogues may, it is argued, increase the perceived legitimacy of the court and by so doing encourage increased compliance with treatment and desistance from crime