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Identification of edgetic mutations that disrupt transcripti

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Simon Chavée

on 5 September 2016

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Transcript of Identification of edgetic mutations that disrupt transcripti

Le complexe Rpd3
The transcriptional regulation
Interactions detection
With yeast genetics by the selection of mutant strains on medium lacking of Uracil :
An essential interaction between Ume6 and URS1
Ume6
An innovative approach
Discussion of results
Test all the possible substitutions
Identification of edgetic mutations that disrupt transcriptional regulation by the yeast Rpd3 histone deacetylase complex
One gene = one protein = one function
Huge improvements and knowledges
Limits
Our comprehension of life, the classical genetics
Systems biology : the whole is greater than the sum of the parts.
The Edgetics : interactions identification with high precision
An holistic approach
By studying the network of macromolecular interactions
With tools of hight technology
Identify macromolecular sequences involved in interactions
With the compansatory mutations
Our study Model :
The Rpd3 complex

Results
Discussion
The hystone deacetylase function
The Rpd3 protein complex
With the next generation sequencing (NGS)
URS1

Use a titrable markers to have a better feeling of the effect of each substitution (lacZ, RT qPCR)
Perspectives
43,2% of isolated mutants Ura+ had a mutation in the URS1
URS1 spontaeus mutations occurs at a frenquency of : 6,5 *10^(-9)
In this context, no substitution of the two last nucleotides
Guanine/Cytosine are often substituted with a Adenine/Thymine and vice versa
Evidences of the important role plays by nucleatide at position 2,3 and 6
Test the regulation of URS1 in other contexts
Start experiments to obtain compensatory mutations
Study other interactors of the Rpd3 complex (Ume6p)
?
A subunits that binds to URS1
Constituvely expressed
Different hypothesis to explain the regulation
A sequence presents in the promoter of many genes
Constituted of a sequence of 10 nucleotides
Thank you for your attention and special thanks to...
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