RNA-RNA and RNA-protein interactions lie at the heart of essential steps of the eukaryotic gene expression pathway. Defects in these processes due to inherited mutations can result in degeneration of the retina, cerebellum, and other tissues. The proposed studies will result in a better understanding of RNA-based mechanisms of gene expression in both the normal and disease states. In particular, we will determine the molecular mechanism for U4/U6.U5 tri-snRNP assembly and activation in pre-mRNA splicing, and for Sen1- dependent termination of transcription by RNA polymerase II. Inherited defects in the first process can lead to autosomal dominant retinitis pigmentosa (adRP), which results in progressive blindness, and in the second process can lead to ataxia ocular apraxia type 2 (AOA2), which results in progressive muscle weakness (cerebellar ataxia). In neither case is the relationship between the molecular defect and the disease understood. Better knowledge of the molecular mechanisms of pre-mRNA splicing and transcription termination should lead to more accurate diagnosis and prognosis of these and other diseases, and may ultimately result in new therapeutic approaches. Furthermore, the proposed studies will illuminate basic mechanisms of eukaryotic gene expression that can be exploited for synthetic biology and biotechnology.

Public Health Relevance

DNA transcription and RNA splicing are essential early steps in gene expression in all human cells. A significant fraction of inherited diseases are due to defects in these two processes, thus the results of our studies could lead to improved disease prediction, diagnosis and therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Unknown (R35)
Project #
3R35GM118075-04S1
Application #
9938191
Study Section
Special Emphasis Panel (ZGM1)
Program Officer
Adkins, Ronald
Project Start
2016-07-01
Project End
2021-06-30
Budget Start
2019-07-01
Budget End
2020-06-30
Support Year
4
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Biochemistry
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Montemayor, Eric J; Didychuk, Allison L; Yake, Allyson D et al. (2018) Architecture of the U6 snRNP reveals specific recognition of 3'-end processed U6 snRNA. Nat Commun 9:1749
Didychuk, Allison L; Butcher, Samuel E; Brow, David A (2018) The life of U6 small nuclear RNA, from cradle to grave. RNA 24:437-460
Didychuk, Allison L; Montemayor, Eric J; Carrocci, Tucker J et al. (2017) Usb1 controls U6 snRNP assembly through evolutionarily divergent cyclic phosphodiesterase activities. Nat Commun 8:497
Chen, Xin; Poorey, Kunal; Carver, Melissa N et al. (2017) Transcriptomes of six mutants in the Sen1 pathway reveal combinatorial control of transcription termination across the Saccharomyces cerevisiae genome. PLoS Genet 13:e1006863
Montemayor, Eric J; Didychuk, Allison L; Liao, Honghong et al. (2017) Structure and conformational plasticity of the U6 small nuclear ribonucleoprotein core. Acta Crystallogr D Struct Biol 73:1-8
Rodgers, Margaret L; Didychuk, Allison L; Butcher, Samuel E et al. (2016) A multi-step model for facilitated unwinding of the yeast U4/U6 RNA duplex. Nucleic Acids Res 44:10912-10928