title: The zinc fingers of YY1 bind single-stranded RNA with low sequence specificity.
abstract: Classical zinc fingers (ZFs) are traditionally considered to act as sequence-specific DNA-binding domains. More recently, classical ZFs have been recognised as potential RNA-binding modules, raising the intriguing possibility that classical-ZF transcription factors are involved in post-transcriptional gene regulation via direct RNA binding. To date, however, only one classical ZF-RNA complex, that involving TFIIIA, has been structurally characterised. Yin Yang-1 (YY1) is a multi-functional transcription factor involved in many regulatory processes, and binds DNA via four classical ZFs. Recent evidence suggests that YY1 also interacts with RNA, but the molecular nature of the interaction remains unknown. In the present work, we directly assess the ability of YY1 to bind RNA using in vitro assays. Systematic Evolution of Ligands by EXponential enrichment (SELEX) was used to identify preferred RNA sequences bound by the YY1 ZFs from a randomised library over multiple rounds of selection. However, a strong motif was not consistently recovered, suggesting that the RNA sequence selectivity of these domains is modest. YY1 ZF residues involved in binding to single-stranded RNA were identified by NMR spectroscopy and found to be largely distinct from the set of residues involved in DNA binding, suggesting that interactions between YY1 and ssRNA constitute a separate mode of nucleic acid binding. Our data are consistent with recent reports that YY1 can bind to RNA in a low-specificity, yet physiologically relevant manner.
authors.affiliation: School of Life and Environmental Sciences, University of Sydney, NSW 2006, Australia.
authors.forename: Dorothy C C
authors.initials: DC
authors.lastname: Wai
authors.affiliation: School of Life and Environmental Sciences, University of Sydney, NSW 2006, Australia.
authors.forename: Manar
authors.initials: M
authors.lastname: Shihab
authors.affiliation: School of Life and Environmental Sciences, University of Sydney, NSW 2006, Australia.
authors.forename: Jason K K
authors.initials: JK
authors.lastname: Low
authors.affiliation: School of Life and Environmental Sciences, University of Sydney, NSW 2006, Australia joel.mackay@sydney.edu.au.
authors.forename: Joel P
authors.initials: JP
authors.lastname: Mackay
mesh_terms: D000595:Amino Acid Sequence; D052157:Aptamers, Nucleotide; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D016296:Mutagenesis; D011485:Protein Binding; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D052156:SELEX Aptamer Technique; D050986:YY1 Transcription Factor / Q000737:chemistry* / Q000378:metabolism; D016335:Zinc Fingers
chemical_list: D052157:Aptamers, Nucleotide; D050986:YY1 Transcription Factor; D012313:RNA; D004247:DNA