Variable telomeric repeat synthesis in Paramecium tetraurelia is consistent with misincorporation by telomerase.
Telomeric DNA at the ends of chromosomes consist of short, tandem repeat sequences. The telomeres of Paramecium tetraurelia are made up of variable repeats, whereas Paramecium caudatum telomeric repeats are largely invariant. To investigate variable repeat synthesis in P. tetraurelia, mutated telomerase RNA genes were expressed in vivo. We demonstrate that the P. caudatum telomerase RNA can participate in telomere synthesis when expressed in the P. tetraurelia macronucleus, despite 24% primary sequence divergence of the RNAs between the two species. De novo telomeric repeats from transformants indicate that P. tetraurelia telomerase fidelity is dramatically affected by template substitutions and that misincorporation at a single templating position is likely to account for the majority of P. tetraurelia telomeric DNA variability. Furthermore, we show that fidelity is not solely a function of the RNA moiety, as the P. caudatum telomerase RNA does not impart high fidelity to the chimeric enzyme.
McCormick-Graham
M
M
Department of Pharmacology, Medical School, University of Minnesota, Minneapolis 55455, USA.
Haynes
W J
WJ
Romero
D P
DP
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000096:biosynthesis*; D017125:Genes, Protozoan; D008845:Microinjections; D008957:Models, Genetic; D009154:Mutation; D016805:Paramecium tetraurelia / Q000235:genetics*; D016053:RNA, Protozoan / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D012091:Repetitive Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA; D019098:Telomerase / Q000235:genetics; D016615:Telomere / Q000235:genetics*; D014170:Transformation, Genetic
D016054:DNA, Protozoan; D016053:RNA, Protozoan; D011993:Recombinant Fusion Proteins; D019098:Telomerase