Post on 14-Aug-2015
Transformation of Human Pirin into Arabidopsis thaliana Plants and Effects on
Seedling Development
Ashley WilliamsWarpeha Lab, Molecular, Cell & Developmental
Program of the Biological Sciences DeptUniversity of Illinois at Chicago
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)
Human vs. AtPirin1
In HumansTranscription co-regulator
Flavonoid metabolismQuercetinase activity
Putative proto-oncogeneCell cycle regulator
In Plants?Transcription co-regulator
Flavonoid metabolismGermination
Role in apoptosis
Seed to Seedlings in At (0-7days) Transcription co-regulator
Flavonoid metabolismQuercetinase activity
GerminationEarly development
Screening…
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?
Human vs. Pirin1 3-D Structure
Arabidopsis PRN1 model (ModBase))
Human PRN1 with inhibitor (3ACL)
Conserved Residues
100% Conserved Residues in
RED
Arabidopsis PRN1 Questions
Do the non-conserved regions play a role in Pirin1 activity?
What is the functional significance of Pirin1 in plant cell physiology?
wt vs. prn1 Mutant Phenotype
wt prn1
72 hours in white light
100μm 100μm
prn1
Courtesy of D. Orozco-Nunnelly (p43)
Experimental Outline
pUC57 Vector with Human Pirin ORF
Observe
Copy Transient vector Select colonies
Select colonies Destination vectorTransform
Select tansformed plants Propagate
Cloning DataHuman 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
To Date Photos
wt transformed with Human Pirin
prn1 mutant transformed with Human Pirin
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!