A critical control point for regulating gene expression in eukaryotic cells is during mRNA transcription by RNA polymerase II (Pol II). Non-coding RNA molecules (ncRNAs) have emerged as a new class of transcriptional regulators in a variety of cellular systems. Mouse B2 RNA and human Alu RNA are two such ncRNAs;they function as repressors of mRNA transcription by binding directly to Pol II in response to heat shock, a widely used model system for studying the cellular stress response. The long term goal of research funded by the parent grant is to understand how mammalian ncRNAs that bind to Pol II control transcription in biologically important and medically relevant experimental systems. The proposed work will complement and extend the goals of the parent grant by identifying and investigating new ncRNA/Pol II complexes in cells. Doing so will test the hypothesis that Pol II contains a high-affinity docking site for ncRNAs, and that diverse ncRNAs bind this site and function as trans-regulators of Pol II transcription in different cellular systems. The proposal hs one Specific Aim: Identify and characterize novel ncRNA/Pol II complexes that form under distinct cellular conditions. New macromolecular complexes containing ncRNAs and Pol II will be found by immunoprecipitating Pol II and sequencing the associated RNAs. Two cellular systems will be used, one focusing on a program of differentiation and the other on cell cycle progression. For selected ncRNAs the percent of each ncRNA bound to Pol II, and how this changes under different conditions and cell types, will be experimentally determined. In addition, the contribution the selected ncRNAs make to the overall programs of differentiation and cell cycle progression in the two cell types will be investigated. These studies will move the fields o transcriptional regulation and functional ncRNAs in new directions. A new class of macromolecular complexes in cells will be defined that have potential to control gene expression through novel mechanisms. Results from the proposed work will provide the platform for future lines of research that will investigate how ncRNAs that complex with Pol II control transcriptional activity during programs of cellular differentiation and cell cycle progression.

Public Health Relevance

Properly controlling gene expression is essential to sustaining life and avoiding many diseases and cancers;mRNA transcription by RNA polymerase II (Pol II) is central to this process. These studies will identify new macromolecular complexes between Pol II and non-coding RNAs that form during specific programs of differentiation and cell cycle progression. Acquiring this knowledge could reveal novel mechanisms of cellular regulation that have the potential to be widespread.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM068414-11S1
Application #
8568673
Study Section
Special Emphasis Panel (ZGM1-CBB-0 (MI))
Program Officer
Sledjeski, Darren D
Project Start
2003-09-19
Project End
2014-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
11
Fiscal Year
2013
Total Cost
$114,375
Indirect Cost
$39,375
Name
University of Colorado at Boulder
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
007431505
City
Boulder
State
CO
Country
United States
Zip Code
80309
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Kugel, Jennifer F; Goodrich, James A (2013) The regulation of mammalian mRNA transcription by lncRNAs: recent discoveries and current concepts. Epigenomics 5:95-102
Kassube, Susanne A; Fang, Jie; Grob, Patricia et al. (2013) Structural insights into transcriptional repression by noncoding RNAs that bind to human Pol II. J Mol Biol 425:3639-48
Wagner, Stacey D; Yakovchuk, Petro; Gilman, Benjamin et al. (2013) RNA polymerase II acts as an RNA-dependent RNA polymerase to extend and destabilize a non-coding RNA. EMBO J 32:781-90
Ponicsan, Steven L; Houel, Stephane; Old, William M et al. (2013) The non-coding B2 RNA binds to the DNA cleft and active-site region of RNA polymerase II. J Mol Biol 425:3625-38
Kugel, Jennifer F; Goodrich, James A (2012) Non-coding RNAs: key regulators of mammalian transcription. Trends Biochem Sci 37:144-51
Tarallo, Valeria; Hirano, Yoshio; Gelfand, Bradley D et al. (2012) DICER1 loss and Alu RNA induce age-related macular degeneration via the NLRP3 inflammasome and MyD88. Cell 149:847-59
Kaneko, Hiroki; Dridi, Sami; Tarallo, Valeria et al. (2011) DICER1 deficit induces Alu RNA toxicity in age-related macular degeneration. Nature 471:325-30
Goodrich, James A; Kugel, Jennifer F (2010) Dampening DNA binding: a common mechanism of transcriptional repression for both ncRNAs and protein domains. RNA Biol 7:305-9
Wagner, Stacey D; Kugel, Jennifer F; Goodrich, James A (2010) TFIIF facilitates dissociation of RNA polymerase II from noncoding RNAs that lack a repression domain. Mol Cell Biol 30:91-7

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