Reconstitution of chromatin in vitro

Abstract

It is now generally believed that DNA methylation is responsible for genomic imprinting in mammals (1). Recent experimental evidence has provided an elegant mechanism for repression of gene expression by DNA methylation. This evidence suggests that proteins that recognize specifically methylated CpGs may contribute to the formation of inactive chromatin (2-5). Nucleosomes are the basic unit of chromatin, consisting of a core of 146 bp of DNA wrapped around a histone octamer (two molecules of each of H2A, H2B, H3, and H4) and a stretch of linker DNA between adjacent nucleosomes. The binding of a fifth histone, known as a linker histone or H1, promotes the packaging of strings of nucleosomes into a 30 nm chromatin fiber (6). Packaging of DNA into nucleosomes and the chromatin fiber greatly restricts the availability of the DNA for nuclear processes such as transcription.