title: Direct non transcriptional role of NF-Y in DNA replication.
abstract: NF-Y is a heterotrimeric transcription factor, which plays a pioneer role in the transcriptional control of promoters containing the CCAAT-box, among which genes involved in cell cycle regulation, apoptosis and DNA damage response. The knock-down of the sequence-specific subunit NF-YA triggers defects in S-phase progression, which lead to apoptotic cell death. Here, we report that NF-Y has a critical function in DNA replication progression, independent from its transcriptional activity. NF-YA colocalizes with early DNA replication factories, its depletion affects the loading of replisome proteins to DNA, among which Cdc45, and delays the passage from early to middle-late S phase. Molecular combing experiments are consistent with a role for NF-Y in the control of fork progression. Finally, we unambiguously demonstrate a direct non-transcriptional role of NF-Y in the overall efficiency of DNA replication, specifically in the DNA elongation process, using a Xenopus cell-free system. Our findings broaden the activity of NF-Y on a DNA metabolism other than transcription, supporting the existence of specific TFs required for proper and efficient DNA replication.
authors.affiliation: Dipartimento di Scienze della Vita, Università di Modena e Reggio Emilia, via Campi 213/D, 41125 Modena, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Benatti
authors.affiliation: Dipartimento di Scienze della Vita, Università di Modena e Reggio Emilia, via Campi 213/D, 41125 Modena, Italy; College of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom.
authors.forename: Silvia
authors.initials: S
authors.lastname: Belluti
authors.affiliation: INSERM, U1016, Institut Cochin, Paris, France; CNRS, UMR8104, Paris, France; Université Paris Descartes, Sorbonne Paris Cité, Paris, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Miotto
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom.
authors.forename: Julia
authors.initials: J
authors.lastname: Neusiedler
authors.affiliation: Dipartimento di Bioscienze, Università degli Studi di Milano, via Celoria 26, 20133 Milano, Italy.
authors.forename: Diletta
authors.initials: D
authors.lastname: Dolfini
authors.affiliation: Institute of Molecular Genetics, CNRS UMR5535 & Université Montpellier, 1919 route de Mende, 34293 Montpellier cedex 5, France.
authors.forename: Marjorie
authors.initials: M
authors.lastname: Drac
authors.affiliation: Dipartimento di Scienze della Vita, Università di Modena e Reggio Emilia, via Campi 213/D, 41125 Modena, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Basile
authors.affiliation: Institute of Molecular Genetics, CNRS UMR5535 & Université Montpellier, 1919 route de Mende, 34293 Montpellier cedex 5, France.
authors.forename: Etienne
authors.initials: E
authors.lastname: Schwob
authors.affiliation: Dipartimento di Bioscienze, Università degli Studi di Milano, via Celoria 26, 20133 Milano, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Mantovani
authors.affiliation: College of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom.
authors.forename: J Julian
authors.initials: JJ
authors.lastname: Blow
authors.affiliation: Dipartimento di Scienze della Vita, Università di Modena e Reggio Emilia, via Campi 213/D, 41125 Modena, Italy. Electronic address: cimbriano@unimo.it.
authors.forename: Carol
authors.initials: C
authors.lastname: Imbriano
mesh_terms: D000818:Animals; D023081:CCAAT-Binding Factor / Q000235:genetics / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D004261:DNA Replication / Q000235:genetics*; D045325:HCT116 Cells; D006801:Humans; D011401:Promoter Regions, Genetic; D016196:S Phase / Q000235:genetics; D061805:Transcription Elongation, Genetic; D014158:Transcription, Genetic; D014982:Xenopus laevis
chemical_list: D023081:CCAAT-Binding Factor; C113571:CDC45 protein, human; D018797:Cell Cycle Proteins; D004247:DNA
keywords: CCAAT-binding factor; DNA replication; NF-Y; Transcription factors; Xenopus cell-free system