title: RNA connectivity requirements between conserved elements in the core of the yeast telomerase RNP.
abstract: Telomerase is a specialized chromosome end-replicating enzyme required for genome duplication in many eukaryotes. An RNA and reverse transcriptase protein subunit comprise its enzymatic core. Telomerase is evolving rapidly, particularly its RNA component. Nevertheless, nearly all telomerase RNAs, including those of H. sapiens and S. cerevisiae, share four conserved structural elements: a core-enclosing helix (CEH), template-boundary element, template, and pseudoknot, in this order along the RNA. It is not clear how these elements coordinate telomerase activity. We find that although rearranging the order of the four conserved elements in the yeast telomerase RNA subunit, TLC1, disrupts activity, the RNA ends can be moved between the template and pseudoknot in vitro and in vivo. However, the ends disrupt activity when inserted between the other structured elements, defining an Area of Required Connectivity (ARC). Within the ARC, we find that only the junction nucleotides between the pseudoknot and CEH are essential. Integrating all of our findings provides a basic map of functional connections in the core of the yeast telomerase RNP and a framework to understand conserved element coordination in telomerase mechanism.
authors.affiliation: Department of Biology, The Johns Hopkins University, Baltimore, MD, USA.
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Mefford
authors.forename: Qundeel
authors.initials: Q
authors.lastname: Rafiq
authors.forename: David C
authors.identifier: 0000000182423493
authors.initials: DC
authors.lastname: Zappulla
mesh_terms: D017124:Conserved Sequence; D012331:RNA, Fungal / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D019098:Telomerase / Q000378:metabolism*
chemical_list: D012331:RNA, Fungal; D012261:Ribonucleoproteins; D019098:Telomerase