title: Effect of the linkers between the zinc fingers in zinc finger protein 809 on gene silencing and nuclear localization.
abstract: Zinc finger protein 809 (ZFP809) belongs to the Kruppel-associated box-containing zinc finger protein (KRAB-ZFP) family and functions in repressing the expression of Moloney murine leukemia virus (MoMLV). ZFP809 binds to the primer-binding site (PBS)located downstream of the MoMLV-long terminal repeat (LTR) and induces epigenetic modifications at integration sites, such as repressive histone modifications and de novo DNA methylation. KRAB-ZFPs contain consensus TGEKP linkers between C2H2 zinc fingers. The phosphorylation of threonine residues within linkers leads to the inactivation of zinc finger binding to target sequences. ZFP809 also contains consensus linkers between zinc fingers. However, the function of ZFP809 linkers remains unknown. In the present study, we constructed ZFP809 proteins containing mutated linkers and examined their ability to silence transgene expression driven by MLV, binding ability to MLV PBS, and cellular localization. The results of the present study revealed that the linkers affected the ability of ZFP809 to silence transgene expression. Furthermore, this effect could be partly attributed to changes in the localization of ZFP809 proteins containing mutated linkers. Further characterization of ZFP809 linkers is required for understanding the functions and features of KRAB-ZFP-containing linkers.
authors.affiliation: Department of Human Genetics, National Center for Child Health and Development, 2-10-1 Okura, Setagaya, Tokyo 157-8535, Japan. Electronic address: ichida-y@ncchd.go.jp.
authors.forename: Yu
authors.initials: Y
authors.lastname: Ichida
authors.affiliation: Department of Human Genetics, National Center for Child Health and Development, 2-10-1 Okura, Setagaya, Tokyo 157-8535, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Utsunomiya
authors.affiliation: Department of Human Genetics, National Center for Child Health and Development, 2-10-1 Okura, Setagaya, Tokyo 157-8535, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Onodera
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D020868:Gene Silencing; D016086:Lentivirus; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics; D009154:Mutation; D019076:Transgenes; D016335:Zinc Fingers
chemical_list: D004268:DNA-Binding Proteins; C541321:ZFP809 protein, mouse
keywords: Kruppel-associated box-containing zinc finger proteins (KRAB-ZFPs); Retroviral vector; Zinc finger protein 809 (ZFP809)