IN VITRO UBIQUITINATION OF AN ARRESTIN-RELATED TRAFFICKING ADAPTOR BY THE SCHIZOSACCHAROMYCES POMBE E3 LIGASE PUB1

The ubiquitination of proteins at the plasma membrane (PM) serves as an endocytosis signal in the downregulation of PM protein levels and cellular responses to their environment. In Saccharomyces cerevisiae, the E3 ubiquitin ligase Rsp5 ubiquitinates its PM targets by forming complexes with arrestin-related trafficking (ART) adaptor proteins that enable Rsp5 to recognize specific sets of PM proteins. The range of specificity for some of the ARTs has been uncovered in yeast models, but the molecular mechanisms by which the ART proteins present substrates to the Rsp5 catalytic domain for ubiquitination remains poorly understood. The ubiquitination of Art1 on lysine residue K486 by Rsp5 has been shown to play a critical role in the Art1-Rsp5 complex’s function, which suggests that this adaptor ubiquitination likely plays a role in stabilization of the Art1-Rsp5 complex or in forming the correct complex structure to enable recognition of substrates. To better understand the mechanism by which Art1 presents PM substrates to Rsp5, and to uncover the functional significance of Art1 ubiquitination, our lab seeks to structurally characterize the Art1-Rsp5 complex. Here I report the purification and biochemical characterization of Pub1 and Any1, the homologous E3-adaptor pair from S. pombe, and an approach to generate the ubiquitinated form of Any1 in vitro for the purposes of structural studies and further biochemical investigation

Cytoplasmic Localization of PCV1-VP3 is a CRM1-Independent Function

The viral protein PCV1-VP3 is of key interest in the field of cancer therapeutic development. PCV1-VP3 exhibits cancer-specific apoptotic capabilities in p53-null cancer cells, but not in human primary cells. PCV1-VP3 was discovered to exhibit subcellular localization to the cytoplasm of transformed cells in a manner that is independent of CRM1 nuclear export. It was also revealed that two regions of the protein exhibit differential localization capabilities when separated, which raises further questions regarding the functional capabilities of each region with respect to cancer "sensing" and apoptosis induction

Profit in a Nonprofit World: A Fundraising Event Guide for FRAXA

The FRAXA Foundation’s annual fundraising depends heavily on event organization and volunteer management. With the goal of improving event efficiency, we analyzed eight nonprofit fundraising events across five organizations to assess necessary time commitments and volunteer requirements, as well as return on investment. From our findings, we crafted an event guide and synthesized recommendations for FRAXA to increase yearly revenue from fundraisers by encouraging community members to self-organize small scale events such as cornhole tournaments, fundraising walks, and game streams

Development and application of an electronic synoptic report for reporting and management of low-dose computed tomography lung cancer screening examination

Abstract Background Interpretation of Low Dose CT scans and protocol driven management of findings is a key aspect of lung cancer screening program performance. Reliable and reproducible methods are needed to communicate radiologists’ interpretation to the screening program or clinicians driving management decision. Methods We performed an audit of a subset of dictated reports from the PANCAN study to assess for omissions. We developed an electronic synoptic reporting tool for radiologists embedded in a clinical documentation system software. The tool was then used for reporting as part of the Alberta Lung Cancer Screening Study and McGill University Health Centre Pilot Lung Cancer Screening Program. Results Fifty reports were audited for completeness. At least one omission was noted in 30 (70%) of reports, with a major omission (missing lobe, size, type of nodule in report or actionable incidental finding in recommendation section of report) in 24 (48%). Details of the reporting template and functionality such as automated nodule cancer risk assessment, Lung-RADS category assignment, auto-generated narrative type report as well as personalize participant results letter is provided. A description of the system’s performance in its application in 2815 CT reports is then summarized. Conclusions We found that narrative type radiologist reports for lung cancer screening CT examinations frequently lacked specific discrete data elements required for management. We demonstrate the successful implementation of a radiology synoptic reporting system for use in lung cancer screening, and the use of this information to drive program management and communications