title: Specific inhibition of NF-Y subunits triggers different cell proliferation defects.
abstract: Regulated gene expression is essential for a proper progression through the cell cycle. The transcription factor NF-Y has a fundamental function in transcriptional regulation of cell cycle genes, particularly of G2/M genes. In order to investigate common and distinct functions of NF-Y subunits in cell cycle regulation, NF-YA, NF-YB and NF-YC have been silenced by shRNAs in HCT116 cells. NF-YA loss led to a delay in S-phase progression, DNA damage and apoptosis: we showed the activation of the replication checkpoint, through the recruitment of Δp53 and of the replication proteins PCNA and Mcm7 to chromatin. Differently, NF-YB depletion impaired cells from exiting G2/M, but did not interfere with S-phase progression. Gene expression analysis of NF-YA and NF-YB inactivated cells highlighted a common set of hit genes, as well as a plethora of uncommon genes, unveiling a different effect of NF-Y subunits loss on NF-Y binding to its target genes. Chromatin extracts and ChIP analysis showed that NF-YA depletion was more effective than NF-YB in hitting NF-Y recruitment to CCAAT-promoters. Our data suggest a critical role of NF-Y expression, highlighting that the lack of the single subunits are differently perceived by the cells, which activate diverse cell cycle blocks and signaling pathways.
authors.affiliation: Dipartimento di Biologia, Università di Modena e Reggio Emilia, via Campi 213/D, 41125 Modena, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Benatti
authors.forename: Diletta
authors.initials: D
authors.lastname: Dolfini
authors.forename: Alessandra
authors.initials: A
authors.lastname: Viganò
authors.forename: Maria
authors.initials: M
authors.lastname: Ravo
authors.forename: Alessandro
authors.initials: A
authors.lastname: Weisz
authors.forename: Carol
authors.initials: C
authors.lastname: Imbriano
mesh_terms: D023081:CCAAT-Binding Factor / Q000037:antagonists & inhibitors*; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004249:DNA Damage; D004261:DNA Replication; D015870:Gene Expression; D020868:Gene Silencing; D006801:Humans; D021122:Protein Subunits / Q000037:antagonists & inhibitors; D016196:S Phase; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D023081:CCAAT-Binding Factor; C492061:NFYA protein, human; C492063:NFYB protein, human; C469358:NFYC protein, human; D021122:Protein Subunits; D016159:Tumor Suppressor Protein p53