Molecular Modeling of the PTF‐J and PTF‐L Heterotrimeric Complexes, 3‐D Printing, Overexpression, Purification and Structural Analysis of the Complexes using Cryo‐Electron Microscopy and Ptf1a gene Expression Patterns During Pancreas Development in Zebrafish

Abstract

Dallas Independent School District biomedical research students study the master regulatory transcription factor heterotrimeric complexes PTF‐J and PTF‐L that are required for the early and late developmental cascade of the pancreas. The PTF‐J and PTF‐L complexes are composed of a common E‐box protein E12/47, Ptf1a. and either RbpJ or RbpL. The PTF‐J complex is required for the early developmental stage of the pancreas and during the secondary transition of pancreas development, RbpJ is replaced by RbpL generating the PTF‐L complex, which is required for the final maturation steps for the development of the exocrine pancreas. In the current studies biomedical research student utilized the known structures for RbpJ, RpbL, the MyoD bHLH domain and E12/47 bHLH domain to generate molecular models bound to the enhancer sequence found in genes regulated by the heterotrimeric complexes during pancreas development. The PTF‐J and PTF‐L enhancer sequence is composed of an E‐box sequence separated by one to three helical turns from a TC‐box sequence. Biomedical research students performed energy minimization of the complexes and generate a model using 3‐D printing technology. Biomedical research students also study the temporal and spatial expression patterns of the PTF‐J and PTF‐L complexes during pancreas development in Zebrafish using immunofluorescence. The overall goals the program are threefold: first, to expose enthusiastic high school students to an authentic hands‐on biomedical research project, second, to solve the structure of the complexes using Cryo‐Electron Microscopy and third to identify the temporal and spatial expression pattern of the Ptf1a protein during pancreas development in Zebrafish.Support or Funding InformationJunior League of DallasThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.