title: The Ubiquitin-like with PHD and Ring Finger Domains 1 (UHRF1)/DNA Methyltransferase 1 (DNMT1) Axis Is a Primary Regulator of Cell Senescence.
abstract: As senescence develops, cells sequentially acquire diverse senescent phenotypes along with simultaneous multistage gene reprogramming. It remains unclear what acts as the key regulator of the collective changes in gene expression at initiation of senescent reprogramming. Here we analyzed time series gene expression profiles obtained in two different senescence models in human diploid fibroblasts: replicative senescence and H2O2-induced senescence. Our results demonstrate that suppression of DNA methyltransferase 1 (DNMT1)-mediated DNA methylation activity was an initial event prior to the display of senescent phenotypes. We identified seven DNMT1-interacting proteins, ubiquitin-like with PHD and ring finger domains 1 (UHRF1), EZH2, CHEK1, SUV39H1, CBX5, PARP1, and HELLS (also known as LSH (lymphoid-specific helicase) 1), as being commonly down-regulated at the same time point as DNMT1 in both senescence models. Knockdown experiments revealed that, among the DNMT1-interacting proteins, only UHRF1 knockdown suppressed DNMT1 transcription. However, UHRF1 overexpression alone did not induce DNMT1 expression, indicating that UHRF1 was essential but not sufficient for DNMT1 transcription. Although UHRF1 knockdown effectively induced senescence, this was significantly attenuated by DNMT1 overexpression, clearly implicating the UHRF1/DNMT1 axis in senescence. Bioinformatics analysis further identified WNT5A as a downstream effector of UHRF1/DNMT1-mediated senescence. Senescence-associated hypomethylation was found at base pairs -1569 to -1363 from the transcription start site of the WNT5A gene in senescent human diploid fibroblasts. As expected, WNT5A overexpression induced senescent phenotypes. Overall, our results indicate that decreased UHRF1 expression is a key initial event in the suppression of DNMT1-mediated DNA methylation and in the consequent induction of senescence via increasing WNT5A expression.
authors.affiliation: From the Departments of Biochemistry and Molecular Biology.|Biomedical Science, Graduate School, Ajou University, Suwon 16499, Korea and.
authors.forename: Hyun-Jung
authors.initials: HJ
authors.lastname: Jung
authors.affiliation: From the Departments of Biochemistry and Molecular Biology.
authors.forename: Hae-Ok
authors.initials: HO
authors.lastname: Byun
authors.affiliation: Biomedical Science, Graduate School, Ajou University, Suwon 16499, Korea and.|Physiology, and.
authors.forename: Byul A
authors.initials: BA
authors.lastname: Jee
authors.affiliation: From the Departments of Biochemistry and Molecular Biology.|Biomedical Science, Graduate School, Ajou University, Suwon 16499, Korea and.
authors.forename: Seongki
authors.initials: S
authors.lastname: Min
authors.affiliation: From the Departments of Biochemistry and Molecular Biology.|Biomedical Science, Graduate School, Ajou University, Suwon 16499, Korea and.
authors.forename: Un-Woo
authors.initials: UW
authors.lastname: Jeoun
authors.affiliation: From the Departments of Biochemistry and Molecular Biology.|Biomedical Science, Graduate School, Ajou University, Suwon 16499, Korea and.
authors.forename: Young-Kyoung
authors.initials: YK
authors.lastname: Lee
authors.affiliation: the Department of Molecular, Cell, and Cancer Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01605.
authors.forename: Yonghak
authors.initials: Y
authors.lastname: Seo
authors.affiliation: Biomedical Science, Graduate School, Ajou University, Suwon 16499, Korea and hg@ajou.ac.kr.|Physiology, and.
authors.forename: Hyun Goo
authors.initials: HG
authors.lastname: Woo
authors.affiliation: From the Departments of Biochemistry and Molecular Biology, ypeace@ajou.ac.kr.|Biomedical Science, Graduate School, Ajou University, Suwon 16499, Korea and.
authors.forename: Gyesoon
authors.initials: G
authors.lastname: Yoon
mesh_terms: D022762:CCAAT-Enhancer-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D016922:Cellular Senescence; D000090266:Chromobox Protein Homolog 5; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics* / Q000378:metabolism*; D019175:DNA Methylation; D005347:Fibroblasts / Q000166:cytology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D000072417:Protein Domains; D034741:RNA, Small Interfering / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases; D000071818:Wnt-5a Protein / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: C000715594:CBX5 protein, human; D022762:CCAAT-Enhancer-Binding Proteins; D006657:Histones; D034741:RNA, Small Interfering; C085389:WNT5A protein, human; D000071818:Wnt-5a Protein; D000090266:Chromobox Protein Homolog 5; D006861:Hydrogen Peroxide; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; C000619380:DNMT1 protein, human; C406364:UHRF1 protein, human; D044767:Ubiquitin-Protein Ligases; D001616:beta-Galactosidase
keywords: DNA methylation; cellular senescence; gene expression; gene regulation; microarray