Abstract

We propose to study the mechanism of genomic imprinting, a genetic process of importance in development, hormonal regulation and tumorigenesis. The programme has been divided into 2 phases according to the guidelines for the Mentored Clinical Scientist Development Award and will be completed over 5 years. The didactic element of the first phase will include laboratory meetings, research conferences, journal clubs and seminars in the Department of Genetics, as well as formal course work on the molecular genetics of eukaryotes and the nucleus and cell cycle. The research component of this proposal is a test of the hypothesis that matrix-attachment regions (MARs) are involved in genomic imprinting. MARs are DNA sequences which anchor chromatin to a fixed nuclear matrix. Many nuclear factors (such as transcriptional and replication enzymes) are associated with the nuclear matrix, which may explain the many effects in cis found for MARs. As these effects include transcriptional enhancement, DNA replication origination, methylation and chromatin structure determination, they are candidates for controlling genomic imprinting. This is because imprinted domains are characterized by differences in gene expression, replication timing, methylation and chromatin structure between homologous chromosomes. We propose that functional differences in MARs between homologous imprinted regions determine these heterogenous manifestations of imprinting. We describe two MARs from the imprinted domain of distal mouse chromosome 7, the first described from an imprinted region. Supporting our hypothesis that they are important regulatory elements, they exhibit striking sequence conservation with rat. Phase I of this proposal combines the didactic component described with the comprehensive mapping of MARs from this domain. Phase II will concentrate on the functional testing of the MARs by transfection into cultured cells of appropriate constructs, laying the foundation for the production of transgenic mice to fully test the role of these elements in genomic imprinting.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08DK002467-04
Application #
6124725
Study Section
Special Emphasis Panel (SRC)
Program Officer
Hyde, James F
Project Start
1996-12-01
Project End
2001-11-30
Budget Start
1999-12-01
Budget End
2000-11-30
Support Year
4
Fiscal Year
2000
Total Cost
$127,305
Indirect Cost

  Institution

Name
Yale University
Department
Genetics
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520

  Publications

State, Matthew W; Greally, John M; Cuker, Adam et al. (2003) Epigenetic abnormalities associated with a chromosome 18(q21-q22) inversion and a Gilles de la Tourette syndrome phenotype. Proc Natl Acad Sci U S A 100:4684-9
Chai, J-H; Locke, D P; Greally, J M et al. (2003) Identification of four highly conserved genes between breakpoint hotspots BP1 and BP2 of the Prader-Willi/Angelman syndromes deletion region that have undergone evolutionary transposition mediated by flanking duplicons. Am J Hum Genet 73:898-925
Kieffer, Lynda J; Greally, John M; Landres, Inna et al. (2002) Identification of a candidate regulatory region in the human CD8 gene complex by colocalization of DNase I hypersensitive sites and matrix attachment regions which bind SATB1 and GATA-3. J Immunol 168:3915-22
Bayarsaihan, Dashzeveg; Dunai, Judit; Greally, John M et al. (2002) Genomic organization of the genes Gtf2ird1, Gtf2i, and Ncf1 at the mouse chromosome 5 region syntenic to the human chromosome 7q11.23 Williams syndrome critical region. Genomics 79:137-43
Greally, John M (2002) Short interspersed transposable elements (SINEs) are excluded from imprinted regions in the human genome. Proc Natl Acad Sci U S A 99:327-32
Chai, J H; Locke, D P; Ohta, T et al. (2001) Retrotransposed genes such as Frat3 in the mouse Chromosome 7C Prader-Willi syndrome region acquire the imprinted status of their insertion site. Mamm Genome 12:813-21
Henegariu, O; Dunai, J; Chen, X N et al. (2001) A triple color FISH technique for mouse chromosome identification. Mamm Genome 12:462-5
Alami, R; Greally, J M; Tanimoto, K et al. (2000) Beta-globin YAC transgenes exhibit uniform expression levels but position effect variegation in mice. Hum Mol Genet 9:631-6
Greally, J M; Gray, T A; Gabriel, J M et al. (1999) Conserved characteristics of heterochromatin-forming DNA at the 15q11-q13 imprinting center. Proc Natl Acad Sci U S A 96:14430-5