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ChIC and ChEC; genomic mapping of chromatin proteins.

Abstract：

:To map the genomic interaction sites of chromatin proteins, two related methods were developed and experimentally explored in Saccharomyces cerevisiae. The ChIC method (chromatin immunocleavage) consists of tethering a fusion protein (pA-MN) consisting of micrococcal nuclease (MN) and staphylococcal protein A to specifically bound antibodies. The nuclease is kept inactive during the tethering process (no Ca2+). The ChEC method (chromatin endogenous cleavage) consists of expressing fusion proteins in vivo, where MN is C-terminally fused to the proteins of interest. The specifically tethered nucleases are activated with Ca2+ ions to locally introduce double-stranded DNA breaks. We demonstrate that ChIC and ChEC map proteins with a 100-200 bp resolution and excellent specificity. One version of the method is applicable to formaldehyde-fixed nuclei, another to native cells with comparable results. Among various model experiments, these methods were used to address the conformation of yeast telomeres.

journal_name

Mol Cell

journal_title

Molecular cell

authors

Schmid M,Durussel T,Laemmli UK

doi

10.1016/j.molcel.2004.09.007

subject

Has Abstract

pub_date

2004-10-08 00:00:00

pages

147-57

issue

1

eissn

1097-2765

issn

1097-4164

pii

S1097-2765(04)00540-4

journal_volume

16

pub_type

杂志文章

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