In mammals, the most abundant internal RNA modification is covalent linkage of a methyl group to the RNA adenine N6 position (N6-methyladenosine or m6A). Functionality of this modification is evidenced by findings that mice engineered to lack m6A de-methylases display abnormalities in energy expenditure and fertility. We reported that m6A methyltransferases are required to maintain mouse embryonic stem cells at their ground state. However, how m6A RNA methylation exerts these functions remains unclear. Recent evidence from our work and others suggests that a major functional mechanism of m6A RNA methylation is to destabilize RNA. In this proposal, we will test this hypothesis by characterizing newly discovered mammalian m6A methyltransferases (Aim 1), identifying the RNA sequence or structure that imparts methylation specificity (Aim 2), and determining whether m6A methylation regulates RNA stability by interacting with the human antigen R - microRNA regulatory pathway (Aim 3). We believe that these are pioneering studies in identifying molecular mechanisms underlying m6A RNA methylation in mammalian cells at the molecular and cellular levels. Fundamental RNA regulatory mechanisms revealed may create a new paradigm for eukaryotic gene regulation and stimulate studies of other modifications of coding or noncoding RNAs. Furthermore, our results could provide strategies to manipulate stability of specific RNAs to maintain pluripotent stem cells in culture as an unlimited source of material for cell therapy.

Public Health Relevance

The proposed work will elucidate mechanisms underlying RNA methylation, a recently characterized RNA regulatory mechanism, in embryonic stem cells. Our findings could be used to enhance the regenerative capacity of pluripotent stem cells, such as patient-derived induced pluripotent stem cells, for use as cell-based therapies for a variety of diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM110090-04
Application #
9405014
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Bender, Michael T
Project Start
2015-01-01
Project End
2019-12-31
Budget Start
2018-01-01
Budget End
2018-12-31
Support Year
4
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Sanford Burnham Prebys Medical Discovery Institute
Department
Type
DUNS #
020520466
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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