The elongation phase of transcription by RNA polymerase II (Pol II) is highly regulated and intricately integrated into mRNA biosynthesis. In an effort to understand the factors involved and their mechanisms of action, studies are proposed that will help understand the role of P-TEFb and TTF2 in controlling transcription elongation and to what extent RNA processing is functionally coupled to transcription. I. P-TEFb plays a key role in regulating the critical checkpoint to productive elongation that occurs shortly after initiation. The kinase activity of P-TEFb is in turn uniquely regulated by reversible association with an RNP which contains 7SK RNA and HEXIM1 or HEXIM2. Parameters affecting the formation and dissociation of the P-TEFb/7SK/HEXIM complex will be examined using methods in vitro and in vivo. Details of the function of P-TEFb on specific genes and globally over all genes will be gathered using nuclear run-on reactions coupled to array methods. II. Transcription is shut off during mitosis and the RNA Pol I and Pol II termination activity of TTF2 is required for mitotic repression of transcription elongation. The role of TTF2 in mitotic transcription repression will be further studied and potential roles in transcription coupled repair, and interphase transcription will be examined. Also the potential role of TFIIF and other elongation factors in controlling TTF2 activity during transcription will be investigated. III. The processing reactions needed to generate functional mRNAs occur in part during elongation and are influenced by the transcription process. Newly developed assays will be used to elucidate the details of the function of the human capping enzyme and the cap methyl transferase in the context of transcription. Finally, using a newly developed in vitro system we will determine the extent to which polyadenylation is functionally coupled to transcription elongation and termination. Most of the studies described are aimed at understanding basic processes controlling gene expression, but P-TEFb plays a major role in HIV gene expression and may be a target for AIDS therapy. Also because a drug in clinical trials for cancer therapy, flavopiridol, targets P-TEFb, the proposed studies are important. ? ?

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Cell Development and Function Integrated Review Group (CDF)
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Tompkins, Laurie
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University of Iowa
Schools of Medicine
Iowa City
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Price, David H (2018) Transient pausing by RNA polymerase II. Proc Natl Acad Sci U S A 115:4810-4812
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
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
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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