title: The polycomb group protein EED varies in its ability to access the nucleus in porcine oocytes and cleavage stage embryos.
abstract: Chromatin-modifying complexes serve essential functions during mammalian embryonic development. Polycomb group proteins EED, SUZ12, and EZH2 have been shown to mediate methylation of the lysine 27 residue of histone protein H3 (H3K27), an epigenetic mark that is linked with transcriptional repression. H3K27 trimethylation has been shown to be present on chromatin in mature porcine oocytes, pronuclear and 2-cell stage embryos, with H3K27 trimethylation decreasing at the 4-cell stage and not detectable in blastocyst stage embryos. The goals of this study were to determine the intracellular localization of the polycomb group protein EED in porcine oocytes and cleavage stage porcine embryos produced by in vitro fertilization and to determine the binding abilities of karyopherin α subtypes toward EED. Our results revealed that EED had a strong nuclear localization in 4-cell and blastocyst stage embryos and a strong perinuclear staining in GV-stage oocytes; EED was not detectable in the nuclei of pronuclear or 2-cell stage embryos. An in vitro binding assay was performed to assess the ability of EED to interact with a series of karyopherin α subtypes; results from this experiment revealed that EED can interact with several karyopherin α subtypes, but with varying degrees of affinity. Together these data indicate that EED displays a dynamic change in intracellular localization in progression from immature oocyte to cleavage stage embryo and that EED possess differing in vitro binding affinities toward individual karyopherin α subtypes, which may in part regulate the nuclear access of EED during this window of development.
authors.affiliation: Department of Animal Sciences, Purdue University, West Lafayette, IN 47907, United States.
authors.forename: Kallie B
authors.initials: KB
authors.lastname: Foust
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Li
authors.forename: Kieun
authors.initials: K
authors.lastname: Park
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Shihong
authors.initials: S
authors.lastname: Liu
authors.forename: Ryan A
authors.initials: RA
authors.lastname: Cabot
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000502:physiology*; D002970:Cleavage Stage, Ovum / Q000502:physiology*; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D009865:Oocytes / Q000502:physiology*; D063151:Polycomb Repressive Complex 2 / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology*; D013552:Swine / Q000502:physiology*; D028901:alpha Karyopherins / Q000145:classification / Q000235:genetics / Q000378:metabolism
chemical_list: D028901:alpha Karyopherins; D063151:Polycomb Repressive Complex 2