Cytosine DNA methylation is an epigenetic mark for gene silencing that is important in many gene regulatory systems including mammalian imprinting, X-chromosome inactivation, and the silencing of transposons and other DNA sequences containing either direct or inverted repeats. Methylation is important in cancer biology, as tumors often show both genome wide demethylation and hypermethylation of specific tumor suppressor genes. We are studying the mechanisms of DNA methylation control in the model plant Arabldopsis, which has methylation systems that have much in common with mammalian systems. In Arabldopsis, the initial establishment of methylation (de novo methylation) requires the activity of DRM2, a homolog of mammalian DnmtS, which is guided to appropriate targets, at least in part, by small interfering RNAs. Once established, DNA methylation is maintained by three different systems for methylation of cytosines in three different sequence contexts, CG, CHG and CHH (asymmetric). CG methylation is maintained by MET1, a homolog of mammalian Dnmtl CHG methylation is maintained by the CMT3 DNA methyltransferase, which is guided to appropriate targets by the methylation of histone H3. CHH methylation is maintained by the persistent activity of DRM2, which is targeted by small interfering RNAs. In this proposal, we plan to further study these DNA methylation control mechanisms. Genetic screens are being used to identify genes required for proper de novo DNA methylation, and we plan to perform a detailed molecular genetic characterization of these genes to understand more about the mechanism by which they contribute to de novo methylation. We have also begun using structural biology, in collaboration with Dinshaw Patel's laboratory, to help in our understanding of individual components of the de novo DNA methylation pathway. We are also applying genomics techniques the study of RNA-directed DNA methylation in order to assist in our understanding the genome wide functions of DNA methylation. These studies should shed new light on DNA methylation systems in Arabldopsis and have implications for related systems in mammals and other eukaryotes.

Public Health Relevance

This research aims at understanding the basic mechanisms that control DNA methylation, which is a chemical modification that occurs on certain genes and prevents these genes from functioning in a particular tissue. When DNA methylation patterns are not properly maintained, this can cause inappropriate regulation of genes and is a major cause of cancer. Thus a further understanding of the DNA methylation may someday lead to methods for correcting DNA methylation patterning defects.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37GM060398-15
Application #
8507941
Study Section
Special Emphasis Panel (NSS)
Program Officer
Carter, Anthony D
Project Start
2000-01-01
Project End
2018-12-31
Budget Start
2014-02-10
Budget End
2014-12-31
Support Year
15
Fiscal Year
2014
Total Cost
$308,000
Indirect Cost
$108,000
Name
University of California Los Angeles
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
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