Proteome Characterization of a Chromatin Locus Using the Proteomics of Isolated Chromatin Segments Approach
The biological functions of given genomic regions are ruled by the local chromatin composition. The Proteomics of Isolated Chromatin segments approach (PICh) is a powerful and unbiased method to analyze the composition of chosen chromatin segments, provided they are abundant (repeated) or that the organism studied has a small genome. PICh can be used to identify novel and unexpected regulatory factors, or when combined with quantitative mass spectrometric approaches, to characterize the function of a defined factor at the chosen locus, by quantifying composition changes at the locus upon removal/addition of that factor.
Key wordsProteomics Chromatin Protein Purification Repeated Sequences Heterochromatin Telomeres
We would like to thank Titia de Lange for the kind gift of the TRF1 antibody.
- 1.Carey MF, Peterson CL, Smale ST (2009) Chromatin immunoprecipitation (ChIP). Cold Spring Harb Protoc 9:pdb prot5279Google Scholar
- 7.Hirsch JD, Eslamizar L, Filanoski BJ, Malekzadeh N, Haugland RP, Beechem JM, Haugland RP (2002) Easily reversible desthiobiotin binding to streptavidin, avidin, and other biotin-binding proteins: uses for protein labeling, detection, and isolation. Anal Biochem 308:343–357CrossRefPubMedGoogle Scholar
- 8.Saksouk N, Barth TK, Ziegler-Birling C, Olava N, Nowak A, Rey E, Mateos-Langerak J, Urbach S, Reik W, Torres-Pedilla ME, Imhof A, Dejardin J, Simboeck E (2014) Redundant mechanisms to form silent chromatin at pericentromeric regions rely on BEND3 and DNA methylation. Mol Cell 56:580–594CrossRefPubMedGoogle Scholar