title: Identification of purple sea urchin telomerase RNA using a next-generation sequencing based approach.
abstract: Telomerase is a ribonucleoprotein (RNP) enzyme essential for telomere maintenance and chromosome stability. While the catalytic telomerase reverse transcriptase (TERT) protein is well conserved across eukaryotes, telomerase RNA (TR) is extensively divergent in size, sequence, and structure. This diversity prohibits TR identification from many important organisms. Here we report a novel approach for TR discovery that combines in vitro TR enrichment from total RNA, next-generation sequencing, and a computational screening pipeline. With this approach, we have successfully identified TR from Strongylocentrotus purpuratus (purple sea urchin) from the phylum Echinodermata. Reconstitution of activity in vitro confirmed that this RNA is an integral component of sea urchin telomerase. Comparative phylogenetic analysis against vertebrate TR sequences revealed that the purple sea urchin TR contains vertebrate-like template-pseudoknot and H/ACA domains. While lacking a vertebrate-like CR4/5 domain, sea urchin TR has a unique central domain critical for telomerase activity. This is the first TR identified from the previously unexplored invertebrate clade and provides the first glimpse of TR evolution in the deuterostome lineage. Moreover, our TR discovery approach is a significant step toward the comprehensive understanding of telomerase RNP evolution.
authors.affiliation: Department of Chemistry & Biochemistry, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Podlevsky
authors.forename: Manja
authors.initials: M
authors.lastname: Marz
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Qi
authors.forename: Steve
authors.initials: S
authors.lastname: Hoffmann
authors.forename: Peter F
authors.initials: PF
authors.lastname: Stadler
authors.forename: Julian J-L
authors.initials: JJ
authors.lastname: Chen
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004789:Enzyme Activation; D057075:Enzyme Assays; D019143:Evolution, Molecular; D015723:Gene Library; D056426:Genetic Loci; D006066:Gonads / Q000166:cytology; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D012313:RNA / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D047270:Strongylocentrotus purpuratus / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D019098:Telomerase / Q000145:classification / Q000235:genetics* / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase
keywords: next-generation sequencing; ribonucleoprotein; telomerase RNA; telomere