A SILAC-Based Screen for Methyl-CpG Binding Proteins Identifies RBP-J as a DNA Methylation and Sequence-Specific Binding Protein
2011

Identifying RBP-J as a DNA Methylation Binding Protein

publication Evidence: moderate

Author Information

Author(s): Bartels Stefanie J. J., Spruijt Cornelia G., Brinkman Arie B., Jansen Pascal W. T. C., Vermeulen Michiel, Stunnenberg Hendrik G.

Primary Institution: Radboud University Nijmegen, Nijmegen, The Netherlands

Hypothesis

Can a SILAC-based approach identify novel methyl-CpG binding proteins?

Conclusion

The study successfully identifies RBP-J as a methylation-dependent binder to DNA, highlighting its role in the methyl-DNA interactome.

Supporting Evidence

  • The study identified several known methyl-CpG binding proteins using a novel SILAC-based approach.
  • RBP-J was found to bind methylated DNA in a sequence-specific manner.
  • Follow-up experiments confirmed the binding of RBP-J to methylated DNA.

Takeaway

Scientists found a new way to see how certain proteins stick to DNA that has special chemical tags, and they discovered a protein called RBP-J that does this in a specific way.

Methodology

The study used a methyl-CpG pull-down assay combined with stable-isotope labeling by amino acids in cell culture (SILAC) to identify methyl-CpG binding proteins.

Limitations

The study may not have identified all potential methyl-CpG binding proteins due to the specific conditions and sequences used.

Digital Object Identifier (DOI)

10.1371/journal.pone.0025884

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