Characterization of cis- and trans-acting elements in the imprinted human SNURF-SNRPN locus.
The imprinted SNRPN locus is a complex transcriptional unit that encodes the SNURF and SmN polypeptides as well as multiple non-coding RNAs. SNRPN is located within the Prader-Willi and Angelman syndrome (PWS/AS) region that contains multiple imprinted genes, which are coordinately regulated by a bipartite imprinting center (IC). The SNRPN 5' region co-localizes with the PWS-IC and contains two DNase I hypersensitive sites, DHS1 at the SNRPN promoter, and DHS2 within intron 1, exclusively on the paternally inherited chromosome. We have examined DHS1 and DHS2 to identify cis- and trans-acting regulatory elements within the endogenous SNRPN 5' region. Analysis of DHS1 by in vivo footprinting and chromatin immunoprecipitation identified allele-specific interaction with multiple regulatory proteins, including NRF-1, which regulates genes involved in mitochondrial and metabolic functions. DHS2 acted as an enhancer of the SNRPN promoter and contained a highly conserved region that showed allele-specific interaction with unphosphorylated RNA polymerase II, YY1, Sp1 and NRF-1, further suggesting a key role for NRF-1 in regulation of the SNRPN locus. We propose that one or more of the regulatory elements identified in this study may also contribute to PWS-IC function.
Rodriguez-Jato
Sara
S
Department of Biochemistry and Molecular Biology, University of Florida College of Medicine Gainesville, FL 32610, USA.
Nicholls
Robert D
RD
Driscoll
Daniel J
DJ
Yang
Thomas P
TP
D024506:5' Flanking Region; D017204:Angelman Syndrome / Q000235:genetics; D001324:Autoantigens; D001483:Base Sequence; D001665:Binding Sites; D018983:DNA Footprinting; D003850:Deoxyribonuclease I / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D018392:Genomic Imprinting; D006657:Histones / Q000378:metabolism; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics*; D011218:Prader-Willi Syndrome / Q000235:genetics; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D020218:Response Elements; D017411:Ribonucleoproteins, Small Nuclear / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism*; D055517:snRNP Core Proteins
D001324:Autoantigens; D006657:Histones; D009687:Nuclear Proteins; D017411:Ribonucleoproteins, Small Nuclear; C526417:SNRPN protein, human; C445239:SNURF protein, human; D014157:Transcription Factors; D055517:snRNP Core Proteins; D012319:RNA Polymerase II; D003850:Deoxyribonuclease I