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Smith-Magenis Syndrome and the Retinoic Acid Induced 1 Gene Undergraduate Research Projects in the Zies Laboratory Department of Biological Sciences, University of Mary Washington, Fredericksburg, VA Dr. Deborah Zies Current Position Research Position RAI1 is a Transcription Factor Associate Professor of Biological Sciences at UMW since 2006 A Post Doctoral Research Fellow Mayo Clinic, Jacksonville, Florida “ PPARg/TGFb in the Intestinal Epithelium” Education Ph.D. in Biochemistry and Molecular Biology University of Florida, Gainesville, Florida “Characterization of the Rabbit HKa2 Gene” M.S. in Biology Tulane University, New Orleans Louisiana “Characterization of Insertion element IS511” Transcription factors are proteins that regulate the expression of other genes. B RAI1 (B) has been shown to regulate the transcription of several genes (C) that have functions that are related to the symptoms of SMS (D). C Currently, there is nothing known about the factors that regulate the expression of RAI1 (A). D B.A. in Biology Rollins College, Winter Park, Florida The two main goals of the Zies laboratory are: 1. To determine the mechanism by which RAI regulates other genes. 2. To determine the mechanism by which RAI1 is regulated. Smith-Magenis Syndrome (SMS) How Does RAI1 Regulate CLOCK? Human Intellectual Disability Disorder that affects approximately 1/ 25,000 births worlds wide. One gene known to be regulated by RAI1 is CLOCK, a major component of our day/night circadian rhythm. One type of evidence for this regulation came from the luciferase reporter gene assay performed in the Zies Laboratory. Symptoms include: • Mild to moderate intellectual disabilities. • Distinctive facial abnormalities • including full cheeks • prominent lower jaw • square-shaped face (Pictured: d, e, f) • Disrupted sleep patterns • Behavioral Disorders including • self-hugging, • Skin picking (Pictured: g), • impulsiveness. • Short fingers and toes (Pictured: h, i) The regulatory region of CLOCK was cloned into the luciferase reporter gene plasmid. As seen in the figure below (first two bars), there was approximately 3.5 fold higher luciferase activity when the CLOCK plasmid was transcribed in the presence of RAI1 (dotted line represents activity in the absence of RAI1). The CLOCK plasmid was then broken into smaller pieces and the new constructs were then tested for luciferase activity when treated alone and also with RAI1. It was found that all fragments contained the RAI1 regulatory region except for deletion 6(+). Critical Region Is Deleted on Chromosome 17 in SMS Patients An ongoing project of the Zies Laboratory is to identify the smallest sequence that contains the RAI1 regulatory region. Identification of this sequence will help identify other genes that are regulated by RAI1. What Regulates RAI1 transcription? • SMS Patients with deletions were divided into groups depending on their deletion type: • A common deletion (pink) • An atypical deletion (green) • a small deletion (blue) • a large deletion (maroon). • Genes that were in the region in which all patients shared the deletion were sequenced in non-deletion patients and all of those patients had a mutation in the RAI1 gene (red box). While we have made significant progress in understanding the mechanism by which RAI1 regulates other genes, we know very little about what regulates the transcription of RAI1. We have purchased five DNA fosmids from the human genome project (boxed sequences). An ongoing project of the Zies laboratory is to isolate the regulatory region for RAI1 from these DNA fosmids and identify specific sequences involved in the regulation of RAI1. Past Lab Members Christine Chapman Tyler Andrew Cox Christine Bax Bachelor of Science: University of Mary Washington, May 2008 Master of Science: University of New Hampshire, December 2011 Current Position: Research Technician, Tufts University Bachelor of Science: University of Mary Washington, May 2009 Master of Science: University of Miami, May 2012 Current position: Science & Policy Specialist, Billfish Foundation Bachelor of Science: University of Mary Washington, May 2011 Current position: Associate Scientist Pharmaceutical Product Development, Inc. Rose Salzberg Melissa Hadley Bachelor of Science: University of Mary Washington, May 2009 Doctor of Pharmacy: Virginia Commonwealth University Anticipated Graduation Date: May 2013 Bachelor of Science: University of Mary Washington, May 2010 Current position: Graduate student, Eastern Virginia Medical School Anticipated Date: May 2013 Rabia Aman Brent Colin Turner Jonathan Hillyard Bachelor of Science: University of Mary Washington, May 2009 Master of Science: Johns Hopkins School of Public Health, May 2012 Current Position: Medical School Student, Medical College of Virginia Bachelor of Science: University of Mary Washington, May 2010 Master of Science: Drexel University College of Medicine, May 2012 Current Position: Osteopathic Medical Student, VCOM Karen Strat Emily Griffith Brittany Abbatiello Andrew Buckner Bachelor of Science: University of Mary Washington, May 2009 Current position: Graduate student, University of Oklahoma Anticipated Graduation Date: May 2015 Bachelor of Science: University of Mary Washington, May 2011 Current position: Technical Intramural Research Award Trainee National Institutes of Health Bachelor of Science: University of Mary Washington, December 2012 Current position: Lab Technician University of Virginia Bachelor of Science: University of Mary Washington, May 20 12 Current Position: Applying to Medical School Bachelor of Science: University of Mary Washington, May 2012 Current position: Graduate Student, Virginia Tech