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GENETIC ASPECTS OF DNA METHYLATION

Award Number: R37GM035690
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 12/01/1985
PERIOD OF PERFORMANCE END DATE: 03/31/2013

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GENETIC ASPECTS OF DNA METHYLATION - We exploit the fungus Neurospora crassa to elucidate the control and function of DNA methylation, an pigenetic process required for X chromosome-inactivation, genomic imprinting and normal development in mammals. In humans, loss of a DNA methyltransferase (MTase) causes ICF syndrome, loss of the methyl- DNA binding protein MeCP2 causes Rett syndrome and abnormal methylation is associatedwith cancer. . Isolation of Neurospora mutants defective in DNA methylation (dim) has led to insights into the control and function of methylation in eukaryotes. For example, our identification of DIM-5 as a histone H3 MTase demonstrated for the first time that histone modifications can control DNA methylation. Our work is helped by valuable resources including: 1. the sequence of the Neurospora genome, 2. efficient methods to knock out genes in Neurospora, S.epitope tagging systems for localization and biochemical studies, 4. antibodies specific for modified histones and other relevant epitopes and 5. powerful analytical methods (e.g., mass spectrometry, chromatin immunoprecipitation and microarray analysis). Most of our effort will go towards identifying and characterizing gene products involved in DNA methylation. We will identify mutants defective in silencing methylated DNA using both forward and reverse genetics. The latter will take advantage of the genome knock-out project, which will disrupt most Neurospora genes within the next few years. We will test all knock-outs for de-repression of our silenced bar gene. We will also employ a biochemical approach to identify protein(s) that trigger de novo methylation based on our identification of sequences that trigger DNA methylation in vivo. In other work, we will elucidate the function, mechanism and interrelationships of histone modifications associated with methylated DNA. We will determine which gene/protein is responsible for each observed histone modification in this fungus and test the phenotypes of mutants unable to perform the modifications. We also will determine the extent to which the histone modifications depend on each other. In addition, we will characterize the Neurospora 'epigenome' in wildtype and mutants using a 'ChlP-on-chip' approach with stock and custom microarrays and will use proteomics to investigate whether critical histone MTases are in protein complexes. Finally, we will follow up our observations suggesting that histone demethylases influence DNA methylation.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2013 ( Subtotal = $0 )
2013	2012	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	000	27	3/21/2013	NON-COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2012 ( Subtotal = $572,014 )
2012	2012	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	000	27	12/1/2011	NON-COMPETING CONTINUATION	$525,074
2012	2012	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	001	27	3/5/2012	NON-COMPETING CONTINUATION	$46,940
														Subtotal = $572,014

Issue Date FY: 2011 ( Subtotal = $570,068 )
2011	2011	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	000	26	11/25/2010	NON-COMPETING CONTINUATION	$533,791
2011	2011	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	001	26	6/15/2011	NON-COMPETING CONTINUATION	$36,277
														Subtotal = $570,068

Issue Date FY: 2010 ( Subtotal = $578,177 )
2010	2010	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	001	25	3/17/2010	NON-COMPETING CONTINUATION	$52,562
2010	2010	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	000	25	11/19/2009	NON-COMPETING CONTINUATION	$525,615
														Subtotal = $578,177

Issue Date FY: 2009 ( Subtotal = $565,000 )
2009	2009	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	001	24	3/26/2009	NON-COMPETING CONTINUATION	$56,500
2009	2009	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	000	24	12/17/2008	NON-COMPETING CONTINUATION	$508,500
														Subtotal = $565,000

Issue Date FY: 2008 ( Subtotal = $550,917 )
2008	2008	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	000	23	11/30/2007	EXTENSION WITH OR WITHOUT FUNDS	$550,917
														Subtotal = $550,917

Issue Date FY: 2007 ( Subtotal = $465,439 )
2007	2007	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	000	22	11/20/2006	NON-COMPETING CONTINUATION	$465,439
														Subtotal = $465,439

Issue Date FY: 2006 ( Subtotal = $465,463 )
2006	2006	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	000	21	11/9/2005	NON-COMPETING CONTINUATION	$465,463
														Subtotal = $465,463

Issue Date FY: 2005 ( Subtotal = $462,865 )
2005	2005	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Biomedical Research and Research Training	000	20	11/12/2004	NON-COMPETING CONTINUATION	$462,865
														Subtotal = $462,865

Issue Date FY: 2004 ( Subtotal = $516,518 )
2004	2004	UNIVERSITY OF OREGON	5219 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Genetics and Developmental Biology Research and Research Training	000	19	11/19/2003	NON-COMPETING CONTINUATION	$516,518
														Subtotal = $516,518

Issue Date FY: 2003 ( Subtotal = $448,815 )
2003	2003	UNIVERSITY OF OREGON	5215 UNIVERSITY OF OREGON	EUGENE	OR	97403	LANE	USA	Genetics and Developmental Biology Research and Research Training	000	18		COMPETING CONTINUATION	$448,815
														Subtotal = $448,815

Grand Total All Awards = $5,195,276

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GENETIC ASPECTS OF DNA METHYLATION

Award Number: R37GM035690

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 12/01/1985

PERIOD OF PERFORMANCE END DATE: 03/31/2013

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