Phylogeny of NF-YA trans-activation splicing isoforms in vertebrate evolution.

NF-Y is a trimeric pioneer Transcription Factor (TF) whose target sequence –the CCAAT box– is present in ~25% of mammalian promoters. We reconstruct the phylogenetic history of the regulatory NF-YA subunit in vertebrates. We find that in addition to the remarkable conservation of the subunits-interaction and DNA-binding parts, the Transcriptional Activation Domain (TAD) is also conserved (>90% identity among bony vertebrates). We infer the phylogeny of the alternatively spliced exon-3 and partial splicing events of exon-7 –7N and 7C– revealing independent clade-specific losses of these regions. These isoforms shape the TAD. Absence of exon-3 in basal deuterostomes, cartilaginous fishes and hagfish, but not in lampreys, suggests that the "short" isoform is primordial, with emergence of exon-3 in chordates. Exon 7N was present in the vertebrate common ancestor, while 7C is a molecular innovation of teleost fishes. RNA-seq analysis in several species confirms expression of all these isoforms. We identify 3 blocks of amino acids in the TAD shared across deuterostomes, yet structural predictions and sequence analyses suggest an evolutionary drive for maintenance of an Intrinsically Disordered Region –IDR– within the TAD. Overall, these data help reconstruct the logic for alternative splicing of this essential eukaryotic TF. • Reconstruction of NF-YA evolution in vertebrates and in other deuterostomes • Conservation of NF-YA Transcriptional Activation Domain (TAD) • Alternative splicing shapes the Q-rich intrinsically disordered TAD • Emergence/loss of alternatively spliced regions in the TAD • Expression of alternatively spliced exons assessed in vertebrates [ABSTRACT FROM AUTHOR]