title: Comparative genomics on MCM8 orthologous genes reveals the transcriptional regulation by transcription factor E2F.
abstract: Minichromosome maintenance protein (MCM) is composed of six structurally related subunits (MCM2-7) and is essential for eukaryotic DNA replication initiation and early stage of elongation process. Recently human and Xenopus MCM8 was identified as a novel member of MCM protein. Here we characterized MCM8 orthologous genes by using bioinformatics. Human MCM8 showed approximately 90%, 90%, 93%, and 79% total-amino acid identity with mouse, rat, dog, and chicken MCM8, respectively. Human, mouse, rat, dog, and chicken MCM8 gene, consisting of 19, 18, 17, 18, and 18 exons, was mapped to 20p12.3-13, 2F3, 3q36, 24, and 3, respectively. We identified transcription factor E2F-binding motifs in the vicinity of the transcription start site among MCM8 orthologous genes. The mammalian but not chicken E2F-binding motif was accompanied by NF-Y binding motif. MCM8 mRNA was upregulated by E2E1 in human culture cells. Chromatin immunoprecipitation (ChIP) demonstrated the direct association of E2F1 and NF-Y with human MCM8 promoter. The promoter activities of human, rat, and chicken MCM8 were demonstrated to be E2F1-dependent. Analysis of human MCM8 promoter constructs showed that an E2F-binding motif in the vicinity of the transcription initiation site is necessary for the transcriptional activation. We also showed that the transcription of human MCM8 is activated by transcription factors E2F1-4, but not by factors E2F5-8.
authors.affiliation: Department of Life Sciences, Graduate School of Agriculture, Meiji University, 1-1-1 Higashimita, Kawasaki, Kanagawa 214-8571, Japan.
authors.forename: Reiko
authors.initials: R
authors.lastname: Hayashi
authors.forename: Yuya
authors.initials: Y
authors.lastname: Goto
authors.forename: Ayaka
authors.initials: A
authors.lastname: Haga
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kobayashi
authors.forename: Ryuji
authors.initials: R
authors.lastname: Ikeda
authors.forename: Kenichi
authors.initials: K
authors.lastname: Yoshida
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics*; D002645:Chickens / Q000235:genetics*; D047369:Chromatin Immunoprecipitation; D002874:Chromosome Mapping; D002890:Chromosomes, Human, Pair 20; D019295:Computational Biology; D017124:Conserved Sequence; D004285:Dogs; D050684:E2F Transcription Factors / Q000235:genetics / Q000378:metabolism*; D005091:Exons; D017930:Genes, Reporter; D023281:Genomics; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D064110:Minichromosome Maintenance Proteins; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D015533:Transcriptional Activation
chemical_list: D018797:Cell Cycle Proteins; D050684:E2F Transcription Factors; D008156:Luciferases; C471623:MCM8 protein, human; D064110:Minichromosome Maintenance Proteins