Emi2-mediated inhibition of E2-substrate ubiquitin transfer by the anaphase-promoting complex/cyclosome through a D-box-independent mechanism.

Vertebrate eggs are arrested at Metaphase II by Emi2, the meiotic anaphase-promoting complex/cyclosome (APC/C) inhibitor. Although the importance of Emi2 during oocyte maturation has been widely recognized and its regulation extensively studied, its mechanism of action remained elusive. Many APC/C inhibitors have been reported to act as pseudosubstrates, inhibiting the APC/C by preventing substrate binding. Here we show that a previously identified zinc-binding region is critical for the function of Emi2, whereas the D-box is largely dispensable. We further demonstrate that instead of acting through a "pseudosubstrate" mechanism as previously hypothesized, Emi2 can inhibit Cdc20-dependent activation of the APC/C substoichiometrically, blocking ubiquitin transfer from the ubiquitin-charged E2 to the substrate. These findings provide a novel mechanism of APC/C inhibition wherein the final step of ubiquitin transfer is targeted and raise the interesting possibility that APC/C is inhibited by Emi2 in a catalytic manner.

Ligation of cell surface GRP78 with antibody directed against the COOH-terminal domain of GRP78 suppresses Ras/MAPK and PI 3-kinase/AKT signaling while promoting caspase activation in human prostate cancer cells.

We have previously shown that treatment of prostate cancer and melanoma cells expressing GRP78 on their cell surface with antibody directed against the COOH-terminal domain of GRP78 upregulates and activates p53 causing decreased cell proliferation and upregulated apoptosis. In this report, we demonstrate that treatment of 1-LN prostate cancer cells with this antibody decreases cell surface expression of GRP78, Akt(Thr308) and Akt(Ser473) kinase activities and reduces phosphorylation of FOXO, and GSK3beta. This treatment also suppresses activation of ERK1/2, p38 MAPK and MKK3/6; however, it upregulates MKK4 activity. JNK, as determined by its phosphorylation state, is subsequently activated, triggering apoptosis. Incubation of cells with antibody reduced levels of anti-apoptotic Bcl-2, while elevating pro-apoptotic BAD, BAX and BAK expression as well as cleaved caspases-3, -7, -8 and -9. Silencing GRP78 or p53 gene expression by RNAi prior to antibody treatment abrogated these effects. We conclude that antibody directed against the COOH-terminal domain of GRP78 may prove useful as a pan suppressor of proliferative/survival signaling in cancer cells expressing GRP78 on their cell surface.

Promoting developmental transcription.

Animal growth and development depend on the precise control of gene expression at the level of transcription. A central role in the regulation of developmental transcription is attributed to transcription factors that bind DNA enhancer elements, which are often located far from gene transcription start sites. Here, we review recent studies that have uncovered significant regulatory functions in developmental transcription for the TFIID basal transcription factors and for the DNA core promoter elements that are located close to transcription start sites.

Inhibition of the anaphase-promoting complex by the Xnf7 ubiquitin ligase.

Degradation of specific protein substrates by the anaphase-promoting complex/cyclosome (APC) is critical for mitotic exit. We have identified the protein Xenopus nuclear factor 7 (Xnf7) as a novel APC inhibitor able to regulate the timing of exit from mitosis. Immunodepletion of Xnf7 from Xenopus laevis egg extracts accelerated the degradation of APC substrates cyclin B1, cyclin B2, and securin upon release from cytostatic factor arrest, whereas excess Xnf7 inhibited APC activity. Interestingly, Xnf7 exhibited intrinsic ubiquitin ligase activity, and this activity was required for APC inhibition. Unlike other reported APC inhibitors, Xnf7 did not associate with Cdc20, but rather bound directly to core subunits of the APC. Furthermore, Xnf7 was required for spindle assembly checkpoint function in egg extracts. These data suggest that Xnf7 is an APC inhibitor able to link spindle status to the APC through direct association with APC core components.

Promoting faculty professionalism: a case-based approach.

INTRODUCTION: Professionalism is a key attribute for health professionals. Yet, it is unknown how much faculty development is directed toward skills and behaviours of faculty professionalism. Faculty professionalism includes boundaries in teacher-student relationships, self-reflection, assuring one's own fitness for duty, and maintaining confidentiality when appropriate. METHODS: For five years, we have incorporated faculty professionalism as a routine agenda item for the monthly Physician Assistant Programme faculty meetings, allowing faculty members to introduce issues they are comfortable sharing or have questions about. We also have case discussions of faculty professionalism within faculty meetings every three months. RESULTS: Faculty professionalism is important in the daily work lives of faculty members and including this as part of routine agendas verifies its importance. A faculty survey showed that a majority look forward to the quarterly faculty professionalism case discussions. These have included attempted influence in the admissions process, student/faculty social boundaries, civic professionalism, students requesting medical advice, and self-disclosure. CONCLUSION: A preventive approach works better than a reactionary approach to faculty missteps in professionalism. Routine discussion of faculty professionalism normalizes the topic and is helpful to both new and experienced faculty members. We recommend incorporation of faculty professionalism as a regular agenda item in faculty meetings.

Mount Carmel in the Commune: Promoting the Holy Land in Central Italy in the 13th and 14th Centuries

The Carmelite friars were the last of the major mendicant orders to be established in Italy. Originally an eremitical order, they arrived from the Holy Land in the 1240s, decades after other mendicant orders, such as the Franciscans and Dominicans, had constructed churches and cultivated patrons in the burgeoning urban centers of central Italy. In a religious market already saturated with friars, the Carmelites distinguished themselves by promoting their Holy Land provenance, eremitical values, and by developing an institutional history claiming to be descendants of the Old Testament prophet Elijah. By the end of the 13th century the order had constructed thriving churches and convents and leveraged itself into a prominent position in the religious community. My dissertation analyzes these early Carmelite churches and convents, as well as the friars’ interactions with patrons, civic governments, and the urban space they occupied. Through three primary case studies – the churches and convents of Pisa, Siena and Florence – I examine the Carmelites’ approach to art, architecture, and urban space as the order transformed its mission from one of solitary prayer to one of active ministry.

My central questions are these: To what degree did the Carmelites’ Holy Land provenance inform the art and architecture they created for their central Italian churches? And to what degree was their visual culture instead a reflection of the mendicant norms of the time?

I have sought to analyze the Carmelites at the institutional level, to determine how the order viewed itself and how it wanted its legacy to develop. I then seek to determine how and if the institutional model was utilized in the artistic and architectural production of the individual convents. The understanding of Carmelite art as a promotional tool for the identity of the order is not a new one, however my work is the first to consider deeply the order’s architectural aspirations. I also consider the order’s relationships with its de facto founding saint, the prophet Elijah, and its patron, the Virgin Mary, in a more comprehensive manner that situates the resultant visual culture into the contemporary theological and historical contexts.

Promoting Decent Work in Global Supply Chains in Latin America and the Caribbean: Key Issues, Good Practices, Lessons Learned and Policy Insights

The report is based on a desk-based review, drawing upon existing studies of global supply chains (GSCs) to examine their impacts and implications for the development of domestic firms, their contribution to productive transformation and structural change and their impacts on the quantity and quality of jobs in the LAC region. It situates the expansion of GSCs in the region within an analytical framework that recognizes both the economic and social upgrading dimensions and the impacts on firms and workers. Special attention is given to the mechanisms for governing the terms and conditions of engagement between firms and between firms and workers in GSCs, with the aim of identifying ways to jointly pursue the goals of raising competitiveness and of promoting productive employment and decent work.