Tetrahymena telomerase ribonucleoprotein RNA-protein interactions.
Telomerase is an enzyme that is essential for the replication and maintenance of chromosomal termini. It is a ribonucleoprotein consisting of a catalytic subunit, one or more associated proteins, and an integral RNA subunit that serves as a template for the synthesisof telomeric repeats. We identified a Tetrahymena telomerase RNA-protein complex by an electrophoretic mobility shift assay, using telomerase partially purified from whole cell extracts and radiolabeled, in vitro transcribed wild-type Tetrahymena telomerase RNA. Complex formation was specific as unlabeled Tetra-hymena telomerase RNA, but not Escherichia coli ribo-somal RNAs, competitively inhibited complex formation. Binding required concentrations of MgCl2of at least 10 mM and occurred over a wide range of potassium glutamate concentrations (20-220 mM). The RNA-protein complex was optimally reconstituted with a 30 degrees C preincubation for </=5 min, prior to electrophoresis. Certain Tetrahymena telomerase RNAs containing deletions of structures and sequences previously predicted to be involved in protein binding were unable to competitively and specifically inhibit complex formation, suggesting a role in protein binding for the deleted residues or structures.
Autexier
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Bloomfield Centre for Research in Aging, Lady Davis Institute for Medical Research,The Sir Mortimer B. Davis-Jewish General Hospital, Montréal, Quebec H3T 1E2, Canada.
Triki
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D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D004926:Escherichia coli / Q000201:enzymology; D016812:Euplotes / Q000201:enzymology; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism; D016053:RNA, Protozoan / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012261:Ribonucleoproteins / Q000737:chemistry / Q000378:metabolism*; D013045:Species Specificity; D019098:Telomerase / Q000737:chemistry / Q000378:metabolism*; D013768:Tetrahymena / Q000201:enzymology* / Q000235:genetics
D046911:Macromolecular Substances; D015800:Protozoan Proteins; D016053:RNA, Protozoan; D012261:Ribonucleoproteins; D019098:Telomerase