aspb midwest presentation 2012

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!