Tracy L. Johnson, MS - US grants

DESCRIPTION (provided by applicant): Eukaryotic gene expression is tightly controlled to maintain proper cellular function. Eukaryotic genes are transcribed by RNA polymerase II to generate fully-processed (capped, polyadenylated, and spliced), mature messenger RNA molecules. Although the cellular machineries that carry out these reactions (RNA synthesis and RNA processing) have typically been studied as biochemically distinct reactions, they are, in fact, temporally and spatially organized to coordinately orchestrate the proper production of a fully-processed mRNA. Misregulation of any of the tightly controlled events in gene expression can have catastrophic consequences for the cell that, ultimately, lead to disease. For example, misregulation (e.g. incorrect splicing, transcriptional down-regulation, etc.) of genes encoding tumor suppressors can and does lead to cancer and a host of other human diseases. Since co-transcriptional splicing occurs in the context of a chromatin template, it is important to understand the functional links between splicing factors and chromatin-modifying enzymes. Using the model organism, Saccharomyces cerevisiae, work described here characterizes unexpected genetic interactions between the histone acetyltransferase, GCN5, within the context of an intact SAGA complex, and the U2 snRNP components MSL1 and LEA1. Furthermore, the gene encoding Gcn5, via its associated HAT activity, is required for cotranscriptional recruitment of Msl1 and Lea1 to pre-mRNAs in vivo. These studies have led to the hypothesis that Gcn5 coordinates pre-mRNA splicing with transcription through its HAT-mediated effects on transcription. To this end, the following specific aims will be undertaken: 1. Characterize the role of Gcn5 mediated histone acetylation in spliceosome assembly. Chromatin immunoprecipitation and biochemical assays will be utilized to map Gcn5 association with DBP2 and ECM33, its interactions with the U2 snRNP components, its acetylation of histones within the genes (and the functional consequence of this acetylation), and its effect on Pol II transcription. 2. Splicing sensitive microarrays will be used to identify genes whose splicing is affected by Gcn5 activity when cells are grown under normal and stress conditions. These introns will be analyzed to identify features that render their splicing Gcn5-dependent. The mechanism by which Gcn5 affects cotranscriptional splicing of these genes will be characterized by, first, mapping Msl1/Lea1 association with the genes. Then, in vivo splicing will be analyzed using the approaches described in Aim 1. PUBLIC HEALTH RELEVANCE: Eukaryotic gene expression is tightly controlled to maintain proper cellular function;misregulation of any of the reactions that allow proper gene expression (such as RNA synthesis and RNA processing) can have catastrophic consequences for the cell that, ultimately, lead to diseases that pose serious public health crises. Understanding the molecular details underlying proper gene expression will allow us to develop tools that may target specific mechanisms in disease progression. Hence, research that elucidates these basic mechanisms of gene expression are an important part of any public health strategy.

? DESCRIPTION (provided by applicant): We will establish a training program for postdoctoral fellows called UCLA Postdocs' Longitudinal Investment in Faculty Training or UPLIFT. This program involves training in innovative active learning pedagogy from UCLA's Preparing Future Faculty (PFF) course and the Center for Education Innovation in the Life Sciences (CEILS). This will be followed by a mentored teaching experience at our partner institution, California State University, Los Angeles (CSULA). CSULA comprises ~70% underrepresented minorities (URMs) in Science Technology, Engineering and Math (STEM). It has low 6-year graduation rates in the Biochemistry and Biology majors. Individual instructors using traditional lecture format currently teach the Introductory Biology and Introduction to Biochemistry courses. Our scholars will team teach these courses with their CSULA mentors and convert them to an active learning format that incorporates numerous flipped lectures in the scholars' and mentors' areas of expertise. UCLA's Higher Education Research Institute (HERI) will formatively evaluate the program using state-of-the-art assessment approaches. The evaluation data will be used to evolve the courses annually with each new round of scholars building upon and improving class activities from the previous round. Our goal is to reach a steady state of 18 scholars to accelerate transition of CSULA courses to active learning. We will test the hypothesis that by incorporating active learning technology, shown in numerous studies to enhance retention of URMs, we will augment CSULA's retention rate and time-to-degree for Biology and Biochemistry majors. UCLA's research training faculty includes 66 outstanding scientists -- 9 are members of the National Academy of Sciences -- chosen for their records of leadership, scholarship, teaching and mentoring. The training faculty cover a wide breadth of research topics employing the newest technologies. All training faculty participate in intramural, disciplin-specific research activities that allow scholars numerous opportunities to present their research and receive feedback from multiple faculty. Recent PhDs with outstanding research credentials and a commitment to teaching will be actively recruited. Stipend support in year 1 derives from the research mentors, while support in years 2-4 will be IRACDA. In year 1, the trainees will perform research and participate in the PFF course and the CEILS workshops at UCLA. In the following years, trainees will continue research, learn the flipped lecture format in UCLA's Life Sciences Core, and teach an increasing number of CSULA class sessions from years 2 to 3. Course preparation will involve mentoring by faculty at the partner institution while the trainees develop active learning modules. The teaching will be closely monitored by the mentors, program directors and HERI using Classroom Observation Protocols for Undergraduate STEM, which specifically measures active learning. In year 4, the focus will be largely in research. The broad-based administrative support for UPLIFT assures a commitment at UCLA and CSULA to the development of a new generation of highly effective faculty trained in active learning and URM retention.