title: Transgenes encompassing dual-promoter CpG islands from the human TBP and HNRPA2B1 loci are resistant to heterochromatin-mediated silencing.
abstract: The genetic elements that are responsible for establishing a transcriptionally competent, open chromatin structure at a region of the genome that consists only of ubiquitously expressed, housekeeping genes are currently unknown. We demonstrate for the first time through functional analysis in stably transfected tissue culture cells that transgenes containing methylation-free CpG islands spanning the dual divergently transcribed promoters from the human TATA binding protein (TBP)-proteasome component-B1 (PSMB1) and heterogeneous nuclear ribonucleoprotein A2/B1 (HNRPA2B1)-heterochromatin protein 1Hs-gamma (chromobox homolog 3, CBX3) gene loci are sufficient to prevent transcriptional silencing and a variegated expression pattern when integrated within centromeric heterochromatin. In addition, only transgene constructs extending over both the HNRPA2B1 and the CBX3 promoters, and not the HNRPA2B1 promoter alone, were able to confer high and stable long-term EGFP reporter gene expression. These observations suggest that methylation-free CpG islands associated with dual, divergently transcribed promoters possess an independent dominant chromatin opening function and may therefore be major determinants in establishing and maintaining a region of open chromatin at housekeeping gene loci.
authors.affiliation: Nuclear Biology Group, Division of Medical and Molecular Genetics, GKT School of Medicine, King's College London, Guy's Campus, 8th Floor Guy's Tower, Guy's Hospital, London Bridge, SE1 9RT, London, UK. michael.antoniou@kcl.ac.uk
authors.forename: Michael
authors.initials: M
authors.lastname: Antoniou
authors.forename: Lee
authors.initials: L
authors.lastname: Harland
authors.forename: Tracey
authors.initials: T
authors.lastname: Mustoe
authors.forename: Steven
authors.initials: S
authors.lastname: Williams
authors.forename: Jolyon
authors.initials: J
authors.lastname: Holdstock
authors.forename: Ernesto
authors.initials: E
authors.lastname: Yague
authors.forename: Tony
authors.initials: T
authors.lastname: Mulcahy
authors.forename: Mark
authors.initials: M
authors.lastname: Griffiths
authors.forename: Sian
authors.initials: S
authors.lastname: Edwards
authors.forename: Panayiotis A
authors.initials: PA
authors.lastname: Ioannou
authors.forename: Andrew
authors.initials: A
authors.lastname: Mountain
authors.forename: Robert
authors.initials: R
authors.lastname: Crombie
mesh_terms: D001483:Base Sequence; D002503:Centromere; D018899:CpG Islands; D020868:Gene Silencing / Q000502:physiology*; D017930:Genes, Reporter; D006570:Heterochromatin / Q000502:physiology*; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D035181:TATA-Box Binding Protein / Q000235:genetics; D019076:Transgenes
chemical_list: D006570:Heterochromatin; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D035181:TATA-Box Binding Protein