The Third Intron of the Interferon Regulatory Factor-8 Is an Initiator of Repressed Chromatin Restricting Its Expression in Non-Immune Cells

Autor: Ben-Zion Levi, Mamduh Khateb, Lior Gepstein, Nitsan Fourier, Ofer Barnea-Yizhar, Aviva Azriel, Hansjörg Hauser, Manabu Nakayama, Sigal Ram, Ulfert Rand, Ekaterina Kovalev
Rok vydání: 2016
Předmět:
0301 basic medicine
Cellular differentiation
lcsh:Medicine
Gene Expression
Mice
Fluorescence Microscopy
Myocytes
Cardiac
Induced pluripotent stem cell
lcsh:Science
Cells
Cultured
Microscopy
Multidisciplinary
Chromosome Biology
Light Microscopy
Cell Differentiation
Genomics
Flow Cytometry
Chromatin
BAC cloning
Interferon Regulatory Factors
Epigenetics
Research Article
Chromatin Immunoprecipitation
Biology
Real-Time Polymerase Chain Reaction
Genome Complexity
Research and Analysis Methods
Cell Line
03 medical and health sciences
Vector cloning
Gene Types
Genetics
Gene silencing
Animals
Humans
Molecular Biology Techniques
Molecular Biology
Reporter gene
Macrophages
lcsh:R
Intron
Biology and Life Sciences
Computational Biology
Cell Biology
Molecular biology
Introns
Mice
Inbred C57BL
030104 developmental biology
RAW 264.7 Cells
Microscopy
Fluorescence
NIH 3T3 Cells
lcsh:Q
IRF8
Chromatin immunoprecipitation
Reporter Genes
Cloning
Developmental Biology
Zdroj: PLoS ONE
PLoS ONE, Vol 11, Iss 6, p e0156812 (2016)
ISSN: 1932-6203
Popis: Interferon Regulatory Factor-8 (IRF-8) serves as a key factor in the hierarchical differentiation towards monocyte/dendritic cell lineages. While much insight has been accumulated into the mechanisms essential for its hematopoietic specific expression, the mode of restricting IRF-8 expression in non-hematopoietic cells is still unknown. Here we show that the repression of IRF-8 expression in restrictive cells is mediated by its 3rd intron. Removal of this intron alleviates the repression of Bacterial Artificial Chromosome (BAC) IRF-8 reporter gene in these cells. Fine deletion analysis points to conserved regions within this intron mediating its restricted expression. Further, the intron alone selectively initiates gene silencing only in expression-restrictive cells. Characterization of this intron's properties points to its role as an initiator of sustainable gene silencing inducing chromatin condensation with suppressive histone modifications. This intronic element cannot silence episomal transgene expression underlining a strict chromatin-dependent silencing mechanism. We validated this chromatin-state specificity of IRF-8 intron upon in-vitro differentiation of induced pluripotent stem cells (iPSCs) into cardiomyocytes. Taken together, the IRF-8 3rd intron is sufficient and necessary to initiate gene silencing in non-hematopoietic cells, highlighting its role as a nucleation core for repressed chromatin during differentiation.
Databáze: OpenAIRE
Externí odkaz: