Abstract

This grant will support research designed to help answer the following questions: (1) What is the molecular mechanism of mammalian X-chromosome inactivation? (2) What is the biological function(s) of 5-methylcytosine in mammalian DNA? The working hypotheses that will guide this work are: (1) 5- Methylcytosine plays a significant role in mammalian gene regulation and cell differentiation, and (2) 5-Methylcytosine is an important component of the X-chromosome inactivation system.
The specific aims are to: (1) identify and study proteins that bind specifically, in vitro and in vivo, to X-linked promoters and X-chromosomal DNA, (2) prepare and study DNA methylated at cytosine residues in CpG doublets by human DNA methyltransferase, (3) study proteins that bind preferentially to enzymatically methylated DNA, and (4) determine the effect of methylation on proteins identified as a result of aim 1.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
5R01AG008196-03
Application #
3119677
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1988-07-01
Project End
1993-06-30
Budget Start
1990-07-01
Budget End
1991-06-30
Support Year
3
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
City of Hope/Beckman Research Institute
Department
Type
DUNS #
City
Duarte
State
CA
Country
United States
Zip Code
91010

  Publications

Smit, A F (1993) Identification of a new, abundant superfamily of mammalian LTR-transposons. Nucleic Acids Res 21:1863-72
Tommasi, S; LeBon, J M; Riggs, A D et al. (1993) Methylation analysis by genomic sequencing of 5' region of mouse Pgk-1 gene and a cautionary note concerning the method. Somat Cell Mol Genet 19:529-41
Pfeifer, G P; Singer-Sam, J; Riggs, A D (1993) Analysis of methylation and chromatin structure. Methods Enzymol 225:567-83
Singer-Sam, J; Riggs, A D (1993) Quantitative analysis of messenger RNA levels: reverse transcription-polymerase chain reaction single nucleotide primer extension assay. Methods Enzymol 225:344-51
Pfeifer, G P; Riggs, A D (1993) Genomic sequencing. Methods Mol Biol 23:169-81
Cooney, C A (1993) Are somatic cells inherently deficient in methylation metabolism? A proposed mechanism for DNA methylation loss, senescence and aging. Growth Dev Aging 57:261-73
Riggs, A D; Pfeifer, G P (1992) X-chromosome inactivation and cell memory. Trends Genet 8:169-74
Singer-Sam, J; Goldstein, L; Dai, A et al. (1992) A potentially critical Hpa II site of the X chromosome-linked PGK1 gene is unmethylated prior to the onset of meiosis of human oogenic cells. Proc Natl Acad Sci U S A 89:1413-7
Schroth, G P; Siino, J S; Cooney, C A et al. (1992) Intrinsically bent DNA flanks both sides of an RNA polymerase I transcription start site. Both regions display novel electrophoretic mobility. J Biol Chem 267:9958-64
Singer-Sam, J; Chapman, V; LeBon, J M et al. (1992) Parental imprinting studied by allele-specific primer extension after PCR: paternal X chromosome-linked genes are transcribed prior to preferential paternal X chromosome inactivation. Proc Natl Acad Sci U S A 89:10469-73

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