Related Experiment Videos
Poly(ADP-ribosyl)ation regulates CTCF-dependent chromatin insulation.
Wenqiang Yu1, Vasudeva Ginjala, Vinod Pant
1Department of Development & Genetics, Evolution Biology Centre, Uppsala University, Norbyvägen 18A, S-752 36 Uppsala, Sweden.
Nature Genetics
|September 14, 2004
Summary
Poly(ADP-ribosyl)ation modifies CTCF, a key chromatin insulator protein. This modification is crucial for CTCF
Area of Science:
- Molecular Biology
- Epigenetics
- Genomics
Background:
- Chromatin insulators maintain gene expression domains by blocking enhancer/silencer activity.
- The CTCF protein is a critical component of chromatin insulators.
- Post-translational modifications regulate protein function in gene regulation.
Purpose of the Study:
- To investigate the post-translational modification of the CTCF protein.
- To determine the role of poly(ADP-ribosyl)ation in CTCF insulator function.
- To explore the implications of CTCF poly(ADP-ribosyl)ation in gene regulation.
Main Methods:
- Chromatin immunoprecipitation (ChIP) assays to detect poly(ADP-ribosyl)ation marks.
- ChIP-on-chip analysis to identify CTCF target sites genome-wide.
- Insulator trap assays to assess insulator function.
- Inhibition of poly(ADP-ribose) polymerase activity using 3-aminobenzamide.
Main Results:
- CTCF was identified as a poly(ADP-ribosyl)ated protein.
- The poly(ADP-ribosyl)ation mark on CTCF is allele-specific at the H19 imprinting control region.
- Poly(ADP-ribosyl)ation is linked to CTCF binding at over 140 genomic sites.
- Insulator function at most CTCF sites is sensitive to poly(ADP-ribose) polymerase inhibition.
Conclusions:
- Poly(ADP-ribosyl)ation is a key post-translational modification that confers chromatin insulator properties to CTCF.
- This modification is important for CTCF-mediated regulation of gene expression domains at both imprinted and nonimprinted loci.
- Dysregulation of poly(ADP-ribosyl)ation may contribute to pathological conditions involving altered gene expression domains.