Abstract
Genomic imprinting is a phenomenon that restricts transcription to predominantly one parental allele. How this transcriptional duality is regulated is poorly understood. Here we perform an RNA interference screen for epigenetic factors involved in paternal allelic silencing at the Kcnq1ot1 imprinted domain in mouse extraembryonic endoderm stem cells. Multiple factors are identified, including nucleoporin 107 (NUP107). To determine NUP107's role and specificity in Kcnq1ot1 imprinted domain regulation, we deplete Nup107, as well as Nup62, Nup98/96 and Nup153. Nup107, Nup62 and Nup153, but not Nup98/96 depletion, reduce Kcnq1ot1 noncoding RNA volume, displace the Kcnq1ot1 domain from the nuclear periphery, reactivate a subset of normally silent paternal alleles in the domain, alter histone modifications with concomitant changes in KMT2A, EZH2 and EHMT2 occupancy, as well as reduce cohesin interactions at the Kcnq1ot1 imprinting control region. Our results establish an important role for specific nucleoporins in mediating Kcnq1ot1 imprinted domain regulation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cell Cycle Proteins / genetics
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Cell Cycle Proteins / metabolism
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Chromosomal Proteins, Non-Histone / genetics
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Chromosomal Proteins, Non-Histone / metabolism
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Cohesins
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Crosses, Genetic
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Embryo, Mammalian
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Endoderm / cytology
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Endoderm / growth & development
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Endoderm / metabolism*
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Enhancer of Zeste Homolog 2 Protein / genetics
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Enhancer of Zeste Homolog 2 Protein / metabolism
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Female
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Gene Expression Regulation, Developmental
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Genomic Imprinting*
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Histone-Lysine N-Methyltransferase / genetics
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Histone-Lysine N-Methyltransferase / metabolism
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Histones / genetics
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Histones / metabolism
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Male
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Mice
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Myeloid-Lymphoid Leukemia Protein / genetics
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Myeloid-Lymphoid Leukemia Protein / metabolism
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Nuclear Pore Complex Proteins / genetics*
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Nuclear Pore Complex Proteins / metabolism
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Potassium Channels, Voltage-Gated / genetics*
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Potassium Channels, Voltage-Gated / metabolism
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Protein Isoforms / genetics
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Protein Isoforms / metabolism
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RNA, Long Noncoding / genetics*
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RNA, Long Noncoding / metabolism
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Stem Cells / cytology
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Stem Cells / metabolism*
Substances
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Cell Cycle Proteins
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Chromosomal Proteins, Non-Histone
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Histones
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KCNQ1OT1 RNA
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Nuclear Pore Complex Proteins
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Potassium Channels, Voltage-Gated
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Protein Isoforms
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RNA, Long Noncoding
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Myeloid-Lymphoid Leukemia Protein
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Enhancer of Zeste Homolog 2 Protein
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Ezh2 protein, mouse
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G9a protein, mouse
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Histone-Lysine N-Methyltransferase
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Kmt2a protein, mouse