ASPB Midwest Presentation 2012

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  1. 1. Ashley Williams Warpeha Lab, Molecular, Cell & Developmental Program of the Biological Sciences Dept University of Illinois at Chicago
  2. 2. General Lab Interests We are interested in the germination to young seedling transition We are interested in abiotic and biotic signals that are mediated by the G-protein alpha subunit GPA1, particularly those mediated by the downstream effector Pirin1 (PRN1)
  3. 3. Human vs. AtPirin1 In Humans Transcription co-regulator Flavonoid metabolism Quercetinase activity Putative proto-oncogene Cell cycle regulator In Plants? Transcription co-regulator Flavonoid metabolism Germination Role in apoptosis Seed to Seedlings in At (0-7days) Transcription co-regulator Flavonoid metabolism Quercetinase activity Germination Early development Screening
  4. 4. Potential signaling pathway involving Pirin1 (PRN1) from seed to seedling Transcription Quercetin GPA1 Phe PRN1 Phenylpropanoids GCR1 (putative G-protein coupled receptor 1) Abiotic and biotic signals CO Receptors?
  5. 5. Human vs. Pirin1 3-D Structure Arabidopsis PRN1 model (ModBase)) Human PRN1 with inhibitor (3ACL)
  6. 6. Conserved Residues 100% Conserved Residues in RED Arabidopsis PRN1 Do the non-conserved regions play a role in Pirin1 activity? What is the functional significance of Pirin1 in plant cell physiology?
  7. 7. wt vs. prn1 Mutant Phenotype wt prn1 72 hours in white light 100m 100m prn1 Courtesy of D. Orozco-Nunnelly (p43)
  8. 8. Experimental Outline pUC57 Vector with Human Pirin ORF Observe Copy Transient vector Select colonies Select colonies Destination vectorTransform Select tansformed plants Propagate
  9. 9. Cloning Data Human Pirin ORF PCR samples 1 kbp 800 bp 1 kbp 800 bp TOPO Cloning samples 1 kbp 800 bp 3PG Vector Samples 1 kbp 800 bp PPG Vector Samples 3PY Vector Samples PPY Vector Samples
  10. 10. To Date Photos wt transformed with Human Pirin prn1 mutant transformed with Human Pirin
  11. 11. Acknowledgements Katherine (Kate) Warpeha (Project Advisor) DurreShahwar Muhammad (Research technician & manager) Danielle Orozco-Nunnelly (Grad student) Xiaona Fan (Grad student) Jennifer Baek (undergraduate researcher and technician) Elise Hwang (visiting undergrad from St Xavier University) Giuliana Biancardi (visiting undergrad from Harold Washington City College of Chicago) Raquel Mezzich (former undergraduate researcher) Molecular, Cell & Developmental biology program in Biological Sciences at UIC UIC Honors College Thank you NSF for support Thanks to ASPB Midwest committee for selecting my abstract!