title: Structural delineation of stem-loop RNA binding by human TAF15 protein.
abstract: Human TATA binding protein associated factor 2 N (TAF15) and Fused in sarcoma (FUS) are nucleic acid binding proteins belonging to the conserved FET family of proteins. They are involved in diverse processes such as pre-mRNA splicing, mRNA transport, and DNA binding. The absence of information regarding the structural mechanism employed by the FET family in recognizing and discriminating their cognate and non-cognate RNA targets has hampered the attainment of consensus on modes of protein-RNA binding for this family. Our study provides a molecular basis of this RNA recognition using a combination of solution-state NMR spectroscopy, calorimetry, docking and molecular dynamics simulation. Analysis of TAF15-RRM solution structure and its binding with stem-loop RNA has yielded conclusive evidence of a non-canonical mode of RNA recognition. Rather than classical stacking interactions that occur across nitrogen bases and aromatic amino acids on ribonucleoprotein sites, moderate-affinity hydrogen bonding network between the nitrogen bases in the stem-loop RNA and a concave face on the RRM surface primarily mediate TAF15-RRM RNA interaction. We have compared the binding affinities across a set of single-stranded RNA oligonucleotides to conclusively establish that RNA binding is dependent upon structural elements in the RNA rather than sequence.
authors.affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, 110 067, New Delhi, India.
authors.forename: Maruthi
authors.initials: M
authors.lastname: Kashyap
authors.affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, 110 067, New Delhi, India.
authors.forename: Akshay Kumar
authors.initials: AK
authors.lastname: Ganguly
authors.affiliation: International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, 110 067, New Delhi, India.
authors.forename: Neel Sarovar
authors.initials: NS
authors.lastname: Bhavesh
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D015870:Gene Expression; D006801:Humans; D006860:Hydrogen Bonding; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D059372:Nucleotide Motifs; D009843:Oligoribonucleotides / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D000067776:RNA Recognition Motif Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D035381:TATA-Binding Protein Associated Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D009843:Oligoribonucleotides; D000067776:RNA Recognition Motif Proteins; D011994:Recombinant Proteins; C102041:TAF15 protein, human; D035381:TATA-Binding Protein Associated Factors; D012313:RNA