title: A widespread distribution of genomic CeMyoD binding sites revealed and cross validated by ChIP-Chip and ChIP-Seq techniques.
abstract: Identifying transcription factor binding sites genome-wide using chromatin immunoprecipitation (ChIP)-based technology is becoming an increasingly important tool in addressing developmental questions. However, technical problems associated with factor abundance and suitable ChIP reagents are common obstacles to these studies in many biological systems. We have used two completely different, widely applicable methods to determine by ChIP the genome-wide binding sites of the master myogenic regulatory transcription factor HLH-1 (CeMyoD) in C. elegans embryos. The two approaches, ChIP-seq and ChIP-chip, yield strongly overlapping results revealing that HLH-1 preferentially binds to promoter regions of genes enriched for E-box sequences (CANNTG), known binding sites for this well-studied class of transcription factors. HLH-1 binding sites were enriched upstream of genes known to be expressed in muscle, consistent with its role as a direct transcriptional regulator. HLH-1 binding was also detected at numerous sites unassociated with muscle gene expression, as has been previously described for its mouse homolog MyoD. These binding sites may reflect several additional functions for HLH-1, including its interactions with one or more co-factors to activate (or repress) gene expression or a role in chromatin organization distinct from direct transcriptional regulation of target genes. Our results also provide a comparison of ChIP methodologies that can overcome limitations commonly encountered in these types of studies while highlighting the complications of assigning in vivo functions to identified target sites.
authors.affiliation: National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Lei
authors.forename: Tetsunari
authors.initials: T
authors.lastname: Fukushige
authors.forename: Wei
authors.initials: W
authors.lastname: Niu
authors.forename: Mihail
authors.initials: M
authors.lastname: Sarov
authors.forename: Valerie
authors.initials: V
authors.lastname: Reinke
authors.forename: Michael
authors.initials: M
authors.lastname: Krause
mesh_terms: D000818:Animals; D001665:Binding Sites; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism*; D002843:Chromatin / Q000737:chemistry; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D016678:Genome; D023281:Genomics / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D009124:Muscle Proteins / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic; D019076:Transgenes
chemical_list: D029742:Caenorhabditis elegans Proteins; D002843:Chromatin; D009124:Muscle Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors; C066645:HLH-1 protein, C elegans; D049452:Green Fluorescent Proteins