We have been interested in the protein CTCF, which we first identified some years ago as having properties of an insulator, blocking interaction between enhancers and promoters when placed between them. We demonstrated that this activity plays an important role in regulating parent of origin allele-specific gene expression at the Igf2/H19 imprinted locus. Work in recent years has shown that a principal mode of action of CTCF is to stabilize interactions between CTCF binding sites on DNA, leading to formation of loop domains. Depending on the geometry of the interactions such loops can either exclude an enhancer leading to insulation, or bring enhancer and promoter closer together, leading to activation. Work in other laboratories has shown that the cohesin complex is associated with CTCF at many of its binding sites, and is essential for stabilizing long range contacts and for insulator activity involving those sites. We have recently shown that the only cohesin subunit that makes direct contact with CTCF is SA2, which is external to the cohesin ring. We have also explored the role of various biochemical modifications in CTCF function. We have recently extended our studies of CTCF function by identifying an interaction between CTCF and the DEAD box helicase, p68. We found that a non-coding RNA, SRA, which forms a complex with p68, is essential for this interaction. We are now investigating the nature of the p68-SRA interaction, and the role of the RNA in insulator activity of CTCF. A genome-wide survey of p68 and SRA binding sites is being carried out. We have also identified other proteins that interact with SRA and may be important for early development and for chromatin structure determination.

Project Start
Project End
Budget Start
Budget End
Support Year
7
Fiscal Year
2013
Total Cost
$528,213
Indirect Cost
City
State
Country
Zip Code
Felsenfeld, Gary (2014) A brief history of epigenetics. Cold Spring Harb Perspect Biol 6:
Sutiwisesak, Rujapope; Kitiyanant, Narisorn; Kotchabhakdi, Naiphinich et al. (2014) Induced pluripotency enables differentiation of human nullipotent embryonal carcinoma cells N2102Ep. Biochim Biophys Acta 1843:2611-2619