The phenomenon of mRNA trans-ligation is a poorly understood process in need of novel experimental techniques and insight. This proposal will study mRNA trans-ligation in human cancer cells that was discovered as a response to treatment with a clinically relevant ribonuclease. The extent of mRNA trans- ligation induced by ribonucleases will be characterized by RNA sequencing techniques. An enzyme hypothesized to be responsible for the observed mRNA ligation will be characterized kinetically and mechanistically. A combination of cellular studies and in vitro characterizations of mRNA ligation will bring to light important details about this uncharacterized physiological process. A fundamental understanding of mRNA trans-ligation is likely to lead to an understanding of various normal physiological and disease processes.

Public Health Relevance

The proposed research will investigate the uncharacterized phenomenon of mRNA trans-ligation. A fundamental understanding of this process will be important to understand the role mRNA trans-ligation plays in both normal physiological and various disease processes. Characterization of the enzyme involved in this process will facilitate the design of inhibitors that specifically target mRNA trans-ligation.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM100681-03
Application #
8607195
Study Section
Special Emphasis Panel (ZRG1-F04B-D (20))
Program Officer
Janes, Daniel E
Project Start
2012-02-01
Project End
2015-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
3
Fiscal Year
2014
Total Cost
$53,942
Indirect Cost
Name
University of Wisconsin Madison
Department
Biochemistry
Type
Schools of Earth Sciences/Natur
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Desai, Kevin K; Bingman, Craig A; Cheng, Chin L et al. (2014) Structure of RNA 3'-phosphate cyclase bound to substrate RNA. RNA 20:1560-6
Desai, Kevin K; Cheng, Chin L; Bingman, Craig A et al. (2014) A tRNA splicing operon: Archease endows RtcB with dual GTP/ATP cofactor specificity and accelerates RNA ligation. Nucleic Acids Res 42:3931-42
Desai, Kevin K; Bingman, Craig A; Phillips Jr, George N et al. (2013) Structures of the noncanonical RNA ligase RtcB reveal the mechanism of histidine guanylylation. Biochemistry 52:2518-25