Telomerase from yeast Saccharomyces cerevisiae is active in vitro as a monomer.
A system for isolation of yeast telomerase via RNA affinity tag in TLC1 RNA was developed. Streptavidin aptamer was inserted at two different positions in TLC1 RNA. Telomerase with TLC1 RNA with one of these inserts is functional in vivo and can be isolated by affinity chromatography in vitro. A telomerase preparation isolated using this technique from a strain producing two distinguishable TLC1 RNA molecules (with and without aptameric insertion) resulted in isolation of active telomerase containing only TLC1 RNA with the aptamer. Our results indicate that yeast telomerase is active in vitro as a monomer.
Shcherbakova
D M
DM
Faculty of Chemistry, Lomonosov Moscow State University, Moscow, 119992, Russia.
Sokolov
K A
KA
Zvereva
M I
MI
Dontsova
O A
OA
D001483:Base Sequence; D008969:Molecular Sequence Data; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D019098:Telomerase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D012331:RNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; D019098:Telomerase