title: Novel Roles and Mechanism for Krüppel-like Factor 16 (KLF16) Regulation of Neurite Outgrowth and Ephrin Receptor A5 (EphA5) Expression in Retinal Ganglion Cells.
abstract: Regenerative medicine holds great promise for the treatment of degenerative retinal disorders. Krüppel-like factors (KLFs) are transcription factors that have recently emerged as key tools in regenerative medicine because some of them can function as epigenetic reprogrammers in stem cell biology. Here, we show that KLF16, one of the least understood members of this family, is a POU4F2 independent transcription factor in retinal ganglion cells (RGCs) as early as embryonic day 15. When overexpressed, KLF16 inhibits RGC neurite outgrowth and enhances RGC growth cone collapse in response to exogenous ephrinA5 ligands. Ephrin/EPH signaling regulates RGC connectivity. The EphA5 promoter contains multiple GC- and GT-rich KLF-binding sites, which, as shown by ChIP-assays, bind KLF16 in vivo In electrophoretic mobility shift assays, KLF16 binds specifically to a single KLF site near the EphA5 transcription start site that is required for KLF16 transactivation. Interestingly, methylation of only six of 98 CpG dinucleotides within the EphA5 promoter blocks its transactivation by KLF16 but enables transactivation by KLF2 and KLF15. These data demonstrate a role for KLF16 in regulation of RGC neurite outgrowth and as a methylation-sensitive transcriptional regulator of EphA5 expression. Together, these data identify differential low level methylation as a novel mechanism for regulating KLF16-mediated EphA5 expression across the retina. Because of the critical role of ephrin/EPH signaling in patterning RGC connectivity, understanding the role of KLFs in regulating neurite outgrowth and Eph receptor expression will be vital for successful restoration of functional vision through optic nerve regenerative therapies.
authors.affiliation: From the Departments of Physiological Optics and Vision Science and.
authors.forename: Jianbo
authors.identifier: 0000-0002-3315-3105
authors.initials: J
authors.lastname: Wang
authors.affiliation: the Byers Eye Institute, School of Medicine, Stanford University, Palo Alto, California 94303, the Shiley Eye Institute, University of California San Diego, La Jolla, California 92093, and.
authors.forename: Joana
authors.identifier: 0000-0002-7699-9901
authors.initials: J
authors.lastname: Galvao
authors.affiliation: From the Departments of Physiological Optics and Vision Science and.
authors.forename: Krista M
authors.initials: KM
authors.lastname: Beach
authors.affiliation: Biology and Biochemistry, University of Houston, Houston, Texas 77204.
authors.forename: Weijia
authors.identifier: 0000-0002-4261-5084
authors.initials: W
authors.lastname: Luo
authors.affiliation: the Laboratory of Epigenetics and Chromatin Dynamics, Gastroenterology Research Unit, Epigenomics Translational Program, Center for Individualized Medicine, Departments of Medicine, Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905.
authors.forename: Raul A
authors.identifier: 0000-0002-1640-6780
authors.initials: RA
authors.lastname: Urrutia
authors.affiliation: the Byers Eye Institute, School of Medicine, Stanford University, Palo Alto, California 94303, the Shiley Eye Institute, University of California San Diego, La Jolla, California 92093, and.
authors.forename: Jeffrey L
authors.identifier: 0000-0002-1390-7360
authors.initials: JL
authors.lastname: Goldberg
authors.affiliation: From the Departments of Physiological Optics and Vision Science and Biology and Biochemistry, University of Houston, Houston, Texas 77204, dotteson@central.uh.edu.
authors.forename: Deborah C
authors.identifier: 0000-0001-6093-6837
authors.initials: DC
authors.lastname: Otteson
mesh_terms: D000818:Animals; D019175:DNA Methylation; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D016501:Neurites / Q000378:metabolism*; D036123:Receptor, EphA5 / Q000096:biosynthesis* / Q000235:genetics; D020218:Response Elements / Q000502:physiology*; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D014158:Transcription, Genetic / Q000502:physiology; D015533:Transcriptional Activation / Q000502:physiology*
chemical_list: C486609:Klf16 protein, mouse; D051741:Kruppel-Like Transcription Factors; D036123:Receptor, EphA5
keywords: DNA methylation; Kruppel-like factor (KLF); chromatin immunoprecipitation (ChIP); gene regulation; mouse; neurite outgrowth; neurodevelopment; promoter; protein expression; retina