Title Information
Title
Yeast two-hybrid screen for new transcriptional silencing proteins
Abstract
DNA codes the blueprint for every living organism, but the integrity of the DNA blueprint is threatened by corrupting transposon sequences. Transposons can proliferate and insert copies of themselves into new positions in DNA. The host cell protects itself against transposon expression and movement by targeting transposons for chemical modifications to DNA and associated histone proteins that silence transcription. Methylation of histone H3 at the lysine 9 position (H3K9me) is a conserved transposon silencing modification in eukaryotes. I am using the laboratory plant Arabidopsis thaliana to understand the structure and function of H3K9 methyltransferase proteins that catalyze the H3K9me modification. Three related H3 K9 methyltransferases—SUVH4, SUVH5, and SUVH6—are important factors in the Arabidopsis transcriptional silencing network. SUVH5 and SUVH6 are of particular interest because they each act preferentially on different transposon targets, but the mechanisms underlying their preferences remain to be elucidated. SUVH5 and SUVH6 have long amino terminal extensions beyond the parts of the protein that mediate histone methylation, including a conserved motif near the amino terminus. We hypothesize that the conserved amino terminal motif provides surfaces that interact with other proteins to help guide SUVH5 and SUVH6 to their preferred target transposons. To investigate this hypothesis, the Bender laboratory previously conducted a yeast two hybrid screen for proteins that can interact with the amino terminus of SUVH5 and SUVH6. This screen recovered a putative mRNA processing protein At4g36980. For my UTRA project, I designed, constructed, and tested mutations in the conserved amino terminal motifs of SUVH5 and SUVH6 to determine whether these mutations disrupt the yeast two hybrid interaction with At4g36980. I also designed, constructed and tested deletions in At4g36980 to determine whether a central serine-arginine (SR) rich region is the critical SUVH interaction motif.
Name: Personal
Name Part
Pallango, Savannah
Role
Role Term
creator
Name: Corporate
Name Part
Brown University. Undergraduate Teaching and Research Awards
Role
Role Term
research program
Name: Personal
Name Part
Bender, Judith.
Role
Role Term
advisor
affiliation
Brown University. Department of Molecular Biology, Cellular Biology and Biochemistry
Origin Information
Publisher
Brown University
Date Created (encoding="w3cdtf")
2014-08-07
Place
Place Term: Text
Providence
Genre (aat)
posters
Subject (Local)
Topic
Yeast two-hybrid
Subject (Local)
Topic
Gene silencing
Subject (Local)
Topic
Transcription
Subject (Local)
Topic
Histone
Subject (Local)
Topic
Methylation
Subject (Local)
Topic
SUVH5
Subject (Local)
Topic
SUVH6
Subject (Local)
Topic
SUVH
Subject (Local)
Topic
Arabidopsis
Subject (Local)
Topic
Transposons
Identifier: DOI
10.26300/p5ct-w290
Type of Resource
text