*Revised Manuscript (unmarked) 1 EVOLUTIONARY HISTORY OF THE PODOPLANIN GENE§ Jaime Renart1*, Diego San Mauro2, Ainhoa Agorreta2, Kim Rutherford3, Neil J. Gemmell3, Miguel Quintanilla1 1Instituto de Investigaciones Biomédicas Alberto Sols, Consejo Superior de Investigaciones Científicas (CSIC)-Universidad Autónoma de Madrid. Spain 2Department of Biodiversity, Ecology, and Evolution. Faculty of Biological Sciences. Universidad Complutense de Madrid. 28040 Madrid. Spain 3Department of Anatomy, School of Biomedical Sciences. University of Otago, PO Box 56, Dunedin 9054, New Zealand *Corresponding author: Jaime Renart Instituto de Investigaciones Biomédicas Alberto Sols, CSIC-UAM Arturo Duperier 4. 28029-Madrid. Spain. T: +34 915854412
[email protected] §We wish to dedicate this publication to the memory of our friend and colleague Luis Álvarez (†2016) 2 Keywords: PDPN, Evolution, Gnathostomes, exon/intron gain Abbreviations: BLAST, Basic Local Alignment Search Tool; CT, cytoplasmic domain; EC, extracellular domain; NCBI, National Center for Biotechnology Information; PDPN, podoplanin; SRA, Sequence Read Archive; TAE, Tris Acetate-EDTA buffer; PCR, polymerase chain reaction; UTR, untranslated region 3 ABSTRACT Podoplanin is a type I small mucin-like protein involved in cell motility. We have identified and studied the podoplanin coding sequence in 201 species of vertebrates, ranging from cartilaginous fishes to mammals. The N-terminal signal peptide is coded by the first exon; the transmembrane and intracellular domains are coded by the third exon (except for the last amino acid, coded in another exon with a long 3’-UTR). The extracellular domain has undergone variation during evolutionary time, having a single exon in cartilaginous fishes, teleosts, coelacanths and lungfishes. In amphibians, this single exon has been split in two, and in amniotes, another exon has been acquired, although it has been secondarily lost in Squamata.