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PMID: 19904620 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Why are we where we are? Understanding replication origins and initiation sites in eukaryotes using ChIP-approaches.

Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology ·Vol. 18 ·No. 1 ·2010-01-00 ·Pages 63-77

Schepers A, Papior P

Abstract

DNA replication initiates from origins of replication following a strict sequential activation programme and a conserved temporal order of activation. The number of replication initiation sites varies between species, according to the complexity of the genomes, with an average spacing of 100,000 bp. In contrast to yeast genomes, the location and definition of origins in mammalian genomes has been elusive. Historically, mammalian replication initiation sites have been mapped in situ by systematically searching specific genomic loci for sites that preferentially initiated DNA replication, potential origins by start-site mapping and autonomously replicating sequence experiments, and potential ORC and pre-replicative complex (pre-RC) sites by chromatin immunoprecipitation (ChIP) using antibodies for pre-RC proteins. In the past decade, ChIP has become an important method for analyzing protein/DNA interactions. Classically, ChIP is combined with Southern blotting or PCR. Recently, whole genome-ChIP methods have been very successful in unicellular eukaryotes to understand molecular mechanisms coordinating replication initiation and its flexibility in response to environmental changes. However, in mammalian systems, ChIP with pre-RC antibodies has often been challenging and genome-wide studies are scarce. In this review, we will appraise the progress that has been made in understanding replication origin organization using immunoprecipitation of the ORC and Mcm2-7 complexes. A special focus will be on the advantages and disadvantages of genome-wide ChIP-technologies and their potential impact on understanding metazoan replicators.

MeSH Terms
Chromatin Immunoprecipitation Eukaryotic Cells Genome Humans Replication Origin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Schepers Aloys
Department of Gene Vectors, Helmholtz Zentrum München-German Research Center for Environmental Health, Marchioninistrasse 25, 81377, München, Germany. schepers@helmholtz-muenchen.de
Papior Peer
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Article Info
Journal
Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology
Abbr.
Chromosome Res
ISSN
1573-6849
DOI
10.1007/s10577-009-9087-1
Email
schepers@helmholtz-muenchen.de
Published
2010-01-00
Pages
63-77
Language
English
Region
Netherlands
NLM ID
9313452
Subset
IM
External Links
PubMed source DOI: 10.1007/s10577-009-9087-1 View journal details
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