title: Novel transcription factor variants through RNA-sequencing: the importance of being "alternative".
abstract: Alternative splicing is a pervasive mechanism of RNA maturation in higher eukaryotes, which increases proteomic diversity and biological complexity. It has a key regulatory role in several physiological and pathological states. The diffusion of Next Generation Sequencing, particularly of RNA-Sequencing, has exponentially empowered the identification of novel transcripts revealing that more than 95% of human genes undergo alternative splicing. The highest rate of alternative splicing occurs in transcription factors encoding genes, mostly in Krüppel-associated box domains of zinc finger proteins. Since these molecules are responsible for gene expression, alternative splicing is a crucial mechanism to "regulate the regulators". Indeed, different transcription factors isoforms may have different or even opposite functions. In this work, through a targeted re-analysis of our previously published RNA-Sequencing datasets, we identified nine novel transcripts in seven transcription factors genes. In silico analysis, combined with RT-PCR, cloning and Sanger sequencing, allowed us to experimentally validate these new variants. Through computational approaches we also predicted their novel structural and functional properties. Our findings indicate that alternative splicing is a major determinant of transcription factor diversity, confirming that accurate analysis of RNA-Sequencing data can reliably lead to the identification of novel transcripts, with potentially new functions.
authors.affiliation: Institute of Genetics and Biophysics "Adriano Buzzati-Traverso", National Research Council, 80131 Naples, Italy. margherita.scarpato@igb.cnr.it.
authors.forename: Margherita
authors.initials: M
authors.lastname: Scarpato
authors.affiliation: Institute of Genetics and Biophysics "Adriano Buzzati-Traverso", National Research Council, 80131 Naples, Italy. antonio.federico@igb.cnr.it.
authors.forename: Antonio
authors.initials: A
authors.lastname: Federico
authors.affiliation: Institute of Genetics and Biophysics "Adriano Buzzati-Traverso", National Research Council, 80131 Naples, Italy; Department of Science and Technology, University of Naples "Parthenope", 80143 Naples, Italy. alfredo.ciccodicola@igb.cnr.it
authors.forename: Alfredo
authors.initials: A
authors.lastname: Ciccodicola
authors.affiliation: Institute of Genetics and Biophysics "Adriano Buzzati-Traverso", National Research Council, 80131 Naples, Italy. valerio.costa@igb.cnr.it.
authors.forename: Valerio
authors.initials: V
authors.lastname: Costa
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics; D040901:Proteomics; D016415:Sequence Alignment; D017423:Sequence Analysis, RNA; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics*
chemical_list: D020033:Protein Isoforms; D014157:Transcription Factors