Our long-term goal is to understand the role and regulation of Mediator in the control of gene expression and cell fate specification within the vertebrate central nervous system. Mediator is a multiprotein interface between gene-specific transcription factors and the eukaryotic RNA polymerase II general transcription machinery. In this capacity, Mediator serves to channel regulatory signals from activator and repressor proteins to affect changes in gene expression programs that control diverse physiological processes, including cell growth and homeostasis, development, and differentiation. MED12, an Xq13-encoded 230 kDa Mediator subunit, plays an essential role in neuronal development through selective regulation of neuronal gene expression. Genetic variation in MED12 has been linked to neuropsychiatric illness and X-linked mental retardation (XLMR). However, the molecular bases by which MED12 controls neuronal differentiation through selective gene regulation and the means by which pathologic sequence alterations impact MED12 function leading to behavioral and cognitive defects remain to be clarified. In this regard, we recently identified a functional interaction between the MED12 interface in Mediator and G9a histone methyltransferase required for epigenetic silencing imposed by the RE1 silencing transcription factor/neuron restrictive silencing factor (REST/NRSF), a key determinant of neuronal fate that suppresses neuronal gene expression in non-neuronal and neural progenitor cells. Notably, we found that missense mutations in MED12 responsible for two XLMR disorders, FG syndrome and Lujan syndrome, disrupt its REST-specific corepressor function, thus linking REST-dependent neuronal gene repression with higher-order cognitive function in humans. Because our recent studies implicate REST-dependent neuronal gene repression in epigenetic restriction of neural progenitor cell differentiation, our findings provide a possible epigenetic perspective to explain the role of MED12 in the etiology of XLMR through altered neuronal development. Thus, we hypothesize that XLMR- associated mutations in MED12 disrupt REST-imposed epigenetic restrictions on neuronal gene expression and neural progenitor cell differentiation. To provide support for this hypothesis, we propose the following aims to decipher the role and pathologic implications of MED12/Mediator in REST-dependent epigenetic suppression of neuronal gene expression and differentiation: (1) Elucidate the mechanistic basis of MED12/Mediator in REST-dependent extra-neuronal gene silencing;(2) Elucidate the role and regulation of MED12/Mediator in REST-dependent suppression of neuronal gene expression and differentiation in neural progenitor cells;(3) Elucidate the impact and mechanism of XLMR-associated mutations in MED12 on REST- dependent suppression of neuronal gene expression and differentiation in neural progenitor cells. We expect these studies to have important human health implications for cell replacement therapy in neurological disease as well as the etiology of XLMR.

Public Health Relevance

We expect these studies to have important implications for cell replacement therapy in neurological disease as well as the etiology of developmental and cognitive defects in humans. First, studies proposed herein to elucidate the mechanism (Aim 1) and regulation (Aim 2) of MED12/Mediator in control of neuronal gene expression and differentiation are expected to break new ground and illuminate fundamental aspects of neural stem cell biology that will be essential to guide prospective cell-based therapeutic approaches to repopulate damaged or diseased areas of the nervous system. Second, studies proposed herein to evaluate the impact of XLMR-associated mutations in MED12 on its basic biochemical properties and functional interactions relevant to neuronal gene repression and neural stem cell differentiation (Aim 3) should reveal new mechanistic insight concerning the etiology of XLMR and possibly identify new avenues for therapeutic or remedial intercession.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Project (R01)
Project #
5R01MH085320-04
Application #
8213456
Study Section
Special Emphasis Panel (ZRG1-MNG-B (01))
Program Officer
Beckel-Mitchener, Andrea C
Project Start
2009-04-07
Project End
2014-01-31
Budget Start
2012-02-01
Budget End
2013-01-31
Support Year
4
Fiscal Year
2012
Total Cost
$367,538
Indirect Cost
$120,038
Name
University of Texas Health Science Center San Antonio
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
800772162
City
San Antonio
State
TX
Country
United States
Zip Code
78229
Kämpjärvi, Kati; Park, Min Ju; Mehine, Miika et al. (2014) Mutations in Exon 1 highlight the role of MED12 in uterine leiomyomas. Hum Mutat 35:1136-41
Turunen, Mikko; Spaeth, Jason M; Keskitalo, Salla et al. (2014) Uterine leiomyoma-linked MED12 mutations disrupt mediator-associated CDK activity. Cell Rep 7:654-60
Vulto-van Silfhout, Anneke T; de Vries, Bert B A; van Bon, Bregje W M et al. (2013) Mutations in MED12 cause X-linked Ohdo syndrome. Am J Hum Genet 92:401-6
Spaeth, Jason M; Kim, Nam Hee; Boyer, Thomas G (2011) Mediator and human disease. Semin Cell Dev Biol 22:776-87
Xu, Xuan; Zhou, Haiying; Boyer, Thomas G (2011) Mediator is a transducer of amyloid-precursor-protein-dependent nuclear signalling. EMBO Rep 12:216-22