Article: article from journal or magazin.
Three Ever Shorter Telomere (EST) genes are dispensable for in vitro yeast telomerase activity.
Proceedings of the National Academy of Sciences of the United States of America
Publication types: Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, P.H.S.
Telomerase is a specialized reverse transcriptase consisting of both RNA and protein components. Previous characterization of yeast telomerase function in vivo identified four EST (for ever shorter telomeres) genes that, when mutated, result in the phenotypes expected for a defect in telomerase. Consistent with this genetic prediction, the EST2 gene has recently been shown to encode the catalytic component of telomerase. Using an in vitro assay, we show here that telomerase activity is present in extracts prepared from yeast strains carrying est1-Delta, est3-Delta, and cdc13-2(est) mutations. Therefore, while these three genes are necessary for telomerase function in vivo, they do not encode components essential for core catalytic activity. When Est2p, the one EST gene product found to be essential for catalytic activity, was immunoprecipitated from extracts, the telomerase RNA subunit was also specifically precipitated, supporting the conclusion that these two components are in a stable complex.
Centrifugation, Density Gradient, Cyclin B/genetics, Cyclin B/metabolism, Fungal Proteins/genetics, Fungal Proteins/metabolism, Genes, Fungal, Mutagenesis, Polymerase Chain Reaction, Proteins/genetics, Proteins/metabolism, Saccharomyces cerevisiae/enzymology, Saccharomyces cerevisiae/genetics, Saccharomyces cerevisiae Proteins, Telomerase/genetics, Telomerase/isolation &, purification, Telomere/genetics, Telomere-Binding Proteins
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