title: Enhanced transgene expression by plasmid-specific recruitment of histone acetyltransferase.
abstract: Histone acetylation is associated with the activation of genes on chromosomes. Transgene expression from plasmid DNA might be increased by the acetylation of histones bound to plasmid DNA. To examine this hypothesis, we employed a positive feedback system, using a fusion protein of the sequence-specific DNA binding domain of yeast GAL4 and the histone acetyltransferase (HAT) domain of mouse CREB-binding protein (GAL4-HAT), in which GAL4-HAT promotes its own expression as well as that of a reporter gene product (luciferase). The activator plasmid DNA carrying the gene encoding GAL4-HAT was introduced into mouse Hepa1-6 cells, together with the reporter plasmid DNA, by lipofection. Significantly increased luciferase expression was observed by the co-introduction of the activator plasmid DNA. Moreover, the acetylation of histones bound to the reporter plasmid DNA was enriched by the activator plasmid DNA. These results indicated that the GAL4-HAT system is useful for enhanced transgene expression.
authors.affiliation: Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan; Graduate School of Science and Engineering, Ehime University, 2-5 Bunkyo-cho, Matsuyama 790-8577, Japan.
authors.forename: Mika
authors.initials: M
authors.lastname: Nishihara
authors.affiliation: Graduate School of Science and Engineering, Ehime University, 2-5 Bunkyo-cho, Matsuyama 790-8577, Japan; Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
authors.forename: Genki N
authors.initials: GN
authors.lastname: Kanda
authors.affiliation: Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Suzuki
authors.affiliation: Graduate School of Science and Engineering, Ehime University, 2-5 Bunkyo-cho, Matsuyama 790-8577, Japan.
authors.forename: Shin'ichiro
authors.initials: S
authors.lastname: Yamakado
authors.affiliation: Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
authors.forename: Hideyoshi
authors.initials: H
authors.lastname: Harashima
authors.affiliation: Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan; Graduate School of Science and Engineering, Ehime University, 2-5 Bunkyo-cho, Matsuyama 790-8577, Japan; Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan. Electronic address: hirokam@hiroshima-u.ac.jp.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kamiya
mesh_terms: D000107:Acetylation; D000818:Animals; D050882:CREB-Binding Protein / Q000737:chemistry; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000737:chemistry; D025461:Feedback, Physiological; D017930:Genes, Reporter / Q000235:genetics; D051548:Histone Acetyltransferases / Q000737:chemistry / Q000378:metabolism*; D006657:Histones / Q000737:chemistry / Q000378:metabolism; D008156:Luciferases / Q000235:genetics; D051379:Mice; D010957:Plasmids / Q000235:genetics*; D000072417:Protein Domains; D011499:Protein Processing, Post-Translational; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry; D014157:Transcription Factors / Q000737:chemistry; D014162:Transfection; D019076:Transgenes / Q000235:genetics*; D015854:Up-Regulation
chemical_list: D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; D006657:Histones; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D008156:Luciferases; D050882:CREB-Binding Protein; C494364:Crebbp protein, mouse; D051548:Histone Acetyltransferases
keywords: Artificial histone acetyltransferase; Chromatin immunoprecipitation; Histone deacetylase inhibitor; Plasmid DNA; Transgene expression