title: Zscan4 is regulated by PI3-kinase and DNA-damaging agents and directly interacts with the transcriptional repressors LSD1 and CtBP2 in mouse embryonic stem cells.
abstract: The Zscan4 family of genes, encoding SCAN-domain and zinc finger-containing proteins, has been implicated in the control of early mammalian embryogenesis as well as the regulation of pluripotency and maintenance of genome integrity in mouse embryonic stem cells. However, many features of this enigmatic family of genes are poorly understood. Here we show that undifferentiated mouse embryonic stem cell (ESC) lines simultaneously express multiple members of the Zscan4 gene family, with Zscan4c, Zscan4f and Zscan4-ps2 consistently being the most abundant. Despite this, between only 0.1 and 0.7% of undifferentiated mouse pluripotent stem cells express Zscan4 protein at a given time, consistent with a very restricted pattern of Zscan4 transcripts reported previously. Herein we demonstrate that Zscan4 expression is regulated by the p110α catalytic isoform of phosphoinositide 3-kinases and is induced following exposure to a sub-class of DNA-damage-inducing agents, including Zeocin and Cisplatin. Furthermore, we observe that Zscan4 protein expression peaks during the G2 phase of the cell cycle, suggesting that it may play a critical role at this checkpoint. Studies with GAL4-fusion proteins suggest a role for Zscan4 in transcriptional regulation, further supported by the fact that protein interaction analyses demonstrate that Zscan4 interacts with both LSD1 and CtBP2 in ESC nuclei. This study advances and extends our understanding of Zscan4 expression, regulation and mechanism of action. Based on our data we propose that Zscan4 may regulate gene transcription in mouse ES cells through interaction with LSD1 and CtBP2.
authors.affiliation: Centre for Regenerative Medicine and Departments of Pharmacy & Pharmacology, University of Bath, Bath, United Kingdom.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Storm
authors.affiliation: Centre for Regenerative Medicine and Departments of Pharmacy & Pharmacology, University of Bath, Bath, United Kingdom; Department of Chemical Engineering, University of Bath, Bath, United Kingdom.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Kumpfmueller
authors.affiliation: Centre for Regenerative Medicine and Departments of Pharmacy & Pharmacology, University of Bath, Bath, United Kingdom.
authors.forename: Heather K
authors.initials: HK
authors.lastname: Bone
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Bath, United Kingdom.
authors.forename: Michael
authors.initials: M
authors.lastname: Buchholz
authors.affiliation: Centre for Regenerative Medicine and Departments of Pharmacy & Pharmacology, University of Bath, Bath, United Kingdom.
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Sanchez Ripoll
authors.affiliation: Department of Chemical Engineering, University of Bath, Bath, United Kingdom.
authors.forename: Julian B
authors.initials: JB
authors.lastname: Chaudhuri
authors.affiliation: RIKEN Centre for Developmental Biology, Kobe, Hyogo, Japan.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Niwa
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Bath, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Tosh
authors.affiliation: Centre for Regenerative Medicine and Departments of Pharmacy & Pharmacology, University of Bath, Bath, United Kingdom.
authors.forename: Melanie J
authors.initials: MJ
authors.lastname: Welham
mesh_terms: D000429:Alcohol Oxidoreductases; D000818:Animals; D001483:Base Sequence; D056970:Co-Repressor Proteins; D004249:DNA Damage; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000378:metabolism*; D056466:Histone Demethylases; D051379:Mice; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D012097:Repressor Proteins / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D000429:Alcohol Oxidoreductases; D056970:Co-Repressor Proteins; D017931:DNA Primers; D004268:DNA-Binding Proteins; D056466:Histone Demethylases; D010089:Oxidoreductases, N-Demethylating; D019869:Phosphatidylinositol 3-Kinases; D010750:Phosphoproteins; D012097:Repressor Proteins; D014157:Transcription Factors; C000588925:Zscan4c protein, mouse; C111341:Ctbp2 protein, mouse; C504076:KDM1a protein, mouse