RNA polymerase II (Pol II) elongation control plays a major role in regulating transcription throughout development and differentiation of multi-cellular organisms. The process is characterized by the default action of negative elongation factors, including NELF and DSIF that halt transcription of initiated polymerases near promoters. These promoter proximal paused polymerases are either released from the template by TTF2 or other termination factors or are allowed to enter productive elongation through the selective action of the positive elongation factor, P-TEFb. The cyclin dependent kinase activity of P-TEFb is regulated by reversible association with the 7SK snRNP mediated by HEXIM proteins. The main goal of this proposal is to further define the mechanisms utilized to control Pol II elongation. The approach will employ biochemical and molecular methods in human cells.
The first aim will explore the role of Gdown1 in the regulation of Pol II elongation. The plan includes in vitro biochemical approaches to purify the Gdown1 negative accessory factor, GNAF, and to purify and characterize a Gdown1 kinase that controls TTF2 dependent termination. In the second aim studies in vitro and in cells will determine mechanistic details of the control of P-TEFb by the 7SK snRNP. The plan describes methods to study the recruitment of P-TEFb to genes to be activated in which the role of the 7SK snRNP will be determined. The regulated release of P- TEFb from the 7SK snRNP by Brd4 and other factors as well as the critical step of re-sequestration of P- TEFb into the 7SK snRNP will be studied. The goal of the final aim is to identify productive elongation factors and determine how they are involved in the transition into and maintenance of productive elongation. The principle investigator is a professor at the University of Iowa and this project wil be carried out in the excellent environment offered by his lab and the facilities in the University of Iowa, Carver College of Medicine. The studies proposed here should significantly increase our understanding of mechanisms controlling gene expression and this will be useful in explaining the disregulation of transcription that occurs in cancer and the hijacking of elongation control machinery by the AIDs virus.

Public Health Relevance

The goal of this project is to understand how gene expression is regulated by controlling RNA polymerase II elongation. Disregulation of components of the elongation control machinery leads to developmental defects, cancer and active HIV infections. Our studies will provide the framework to understand and potentially treat these diseases.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Molecular Genetics A Study Section (MGA)
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Sledjeski, Darren D
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University of Iowa
Schools of Medicine
Iowa City
United States
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Price, David H (2018) Transient pausing by RNA polymerase II. Proc Natl Acad Sci U S A 115:4810-4812
Nilson, Kyle A; Lawson, Christine K; Mullen, Nicholas J et al. (2017) Oxidative stress rapidly stabilizes promoter-proximal paused Pol II across the human genome. Nucleic Acids Res 45:11088-11105
Bosque, Alberto; Nilson, Kyle A; Macedo, Amanda B et al. (2017) Benzotriazoles Reactivate Latent HIV-1 through Inactivation of STAT5 SUMOylation. Cell Rep 18:1324-1334
Mullen, Nicholas J; Price, David H (2017) Hydrogen peroxide yields mechanistic insights into human mRNA capping enzyme function. PLoS One 12:e0186423
Brogie, John E; Price, David H (2017) Reconstitution of a functional 7SK snRNP. Nucleic Acids Res 45:6864-6880
Tan, Justin L; Fogley, Rachel D; Flynn, Ryan A et al. (2016) Stress from Nucleotide Depletion Activates the Transcriptional Regulator HEXIM1 to Suppress Melanoma. Mol Cell 62:34-46
Nilson, Kyle A; Guo, Jiannan; Turek, Michael E et al. (2015) THZ1 Reveals Roles for Cdk7 in Co-transcriptional Capping and Pausing. Mol Cell 59:576-87
Guo, Jiannan; Li, Tiandao; Schipper, Joshua et al. (2014) Sequence specificity incompletely defines the genome-wide occupancy of Myc. Genome Biol 15:482
Fowler, Trent; Ghatak, Payel; Price, David H et al. (2014) Regulation of MYC expression and differential JQ1 sensitivity in cancer cells. PLoS One 9:e87003
Guo, Jiannan; Turek, Michael E; Price, David H (2014) Regulation of RNA polymerase II termination by phosphorylation of Gdown1. J Biol Chem 289:12657-65

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