Format

Send to

Choose Destination
Cell. 2009 Jan 9;136(1):175-86. doi: 10.1016/j.cell.2008.11.045.

Purification of proteins associated with specific genomic Loci.

Author information

1
Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114, USA.

Abstract

Eukaryotic DNA is bound and interpreted by numerous protein complexes in the context of chromatin. A description of the full set of proteins that regulate specific loci is critical to understanding regulation. Here, we describe a protocol called proteomics of isolated chromatin segments (PICh) that addresses this issue. PICh uses a specific nucleic acid probe to isolate genomic DNA with its associated proteins in sufficient quantity and purity to allow identification of the bound proteins. Purification of human telomeric chromatin using PICh identified the majority of known telomeric factors and uncovered a large number of novel associations. We compared proteins found at telomeres maintained by the alternative lengthening of telomeres (ALT) pathway to proteins bound at telomeres maintained by telomerase. We identified and validated several proteins, including orphan nuclear receptors, that specifically bind to ALT telomeres, establishing PICh as a useful tool for characterizing chromatin composition.

PMID:
19135898
PMCID:
PMC3395431
DOI:
10.1016/j.cell.2008.11.045
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center