|
NIDA Proteomics Center
>
Investigators
> Arthur Simen, MD, PhD
Proteomic Analysis Of The
Promoter Regions Of Genes Relevant To Substance Abuse And Dependence
Arthur Simen, MD, PhD, Divisions of Molecular Psychiatry, Aging Research, and
Human Genetics, Yale University
Drug addiction is associated with changes in
chromatin state. Drugs of abuse cause long-lasting changes in the state of
proteins associated with DNA, including changes in transcription factor binding
and changes in post-translational histone modifications, and these changes may
underlie drug addiction. Risk factors for substance abuse, including genetic
polymorphisms and stress also affect chromatin structure and function. Current
methods for the study of DNA-protein interactions are limited to the study of
single DNA regions and single proteins, or single proteins and multiple DNA
regions. Methods for comprehensively studying protein occupancy and
post-translational modifications of particular chromatin regions and the many
proteins associated with these regions would significantly advance our
understanding of genome regulation in addiction. New proteomics methods are
ideally suited to characterizing the chromatin-associated proteome. Studies
conducted to date have characterized alterations in transcription factor binding
caused by a novel promoter variant of SLC6A1 that we discovered by resequencing.
Here we propose to develop this methodology further and extend it to the study
of native chromatin. Briefly, we propose to use tiled, biotin labeled
oligonucleotide libraries to isolate chromatin regions of interest from brain
regions implicated in drug addiction from rodent models of addiction and
vulnerability to drug taking. Protein isolated from these chromatin fragments
will be analyzed by iTraq and MIDAS to quantitatively characterize the spectrum
of proteins associated with the chromatin region and to quantify
post-translational histone modifications. This methodology will allow for a
large improvement in the resolution of chromatin analysis in relation to
addictive behavior and is likely to lead to many novel insights that would be
impossible without the availability of powerful new proteomics tools. |
