Association of the Est1 protein with telomerase activity in yeast

The est1 mutant was previously identified because it is defective in telomere maintenance and displays a senescent phenotype. To see if Est1 might be a component of yeast telomerase, we examined immunoprecipitated Est1. The yeast telomerase RNA Tlc1 specifically coprecipitated with Est1. Furthermore...

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Veröffentlicht in:Proceedings of the National Academy of Sciences - PNAS 1996-04, Vol.93 (7), p.2817-2821
Hauptverfasser: Steiner, Barbara R., Hidaka, Kyoko, Futcher, Bruce
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creator Steiner, Barbara R.
Hidaka, Kyoko
Futcher, Bruce
description The est1 mutant was previously identified because it is defective in telomere maintenance and displays a senescent phenotype. To see if Est1 might be a component of yeast telomerase, we examined immunoprecipitated Est1. The yeast telomerase RNA Tlc1 specifically coprecipitated with Est1. Furthermore, the Est1 immunoprecipitates contained a telomerase-like activity. As expected for yeast telomerase, the activity elongated telomeric primers, it required dGTP and dTTP but not dATP or dCTP, and it was sensitive to RNase A. Further evidence suggesting that the activity was telomerase was obtained from experiments using a TLC1-1 mutant strain, which has a mutant telomerase template containing dG residues. The activity immunoprecipitated from TLC1-1 mutant strains incorporated 32P-labeled dCTP, while activity from TLC1 strains did not. Use of different telomeric primer substrates revealed two distinguishable telomerase-like activities: one was dependent on TLC1, and one was not. The TLC1-independent activity may be due to a second yeast telomerase RNA, or it may be some other kind of activity.
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To see if Est1 might be a component of yeast telomerase, we examined immunoprecipitated Est1. The yeast telomerase RNA Tlc1 specifically coprecipitated with Est1. Furthermore, the Est1 immunoprecipitates contained a telomerase-like activity. As expected for yeast telomerase, the activity elongated telomeric primers, it required dGTP and dTTP but not dATP or dCTP, and it was sensitive to RNase A. Further evidence suggesting that the activity was telomerase was obtained from experiments using a TLC1-1 mutant strain, which has a mutant telomerase template containing dG residues. The activity immunoprecipitated from TLC1-1 mutant strains incorporated 32P-labeled dCTP, while activity from TLC1 strains did not. Use of different telomeric primer substrates revealed two distinguishable telomerase-like activities: one was dependent on TLC1, and one was not. The TLC1-independent activity may be due to a second yeast telomerase RNA, or it may be some other kind of activity.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>8610124</pmid><doi>10.1073/pnas.93.7.2817</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record>
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ispartof Proceedings of the National Academy of Sciences - PNAS, 1996-04, Vol.93 (7), p.2817-2821
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subjects actividad enzimatica
activite enzymatique
adn
Amino Acid Sequence
Base Sequence
Biochemistry
Blotting, Northern
Cellular senescence
Centrifugation
chromosome
chromosomes
cromosomas
Deoxyguanosine
dna
Enzymes
enzymic activity
Fungal Proteins - isolation & purification
Fungal Proteins - metabolism
Kinetics
Molecular Sequence Data
Mutagenesis
mutant
mutantes
mutants
Mutation
nucleotide
Nucleotides
nucleotidos
Oligodeoxyribonucleotides - chemistry
Oligodeoxyribonucleotides - metabolism
Oligonucleotides
Phenotypes
pirimidinas
Plasmids
Proteins
purificacion
purification
pyrimidine
pyrimidines
RNA
RNA, Fungal - biosynthesis
RNA, Fungal - isolation & purification
RNA, Fungal - metabolism
Saccharomyces cerevisiae
Saccharomyces cerevisiae - enzymology
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae Proteins
Substrate Specificity
Telomerase - isolation & purification
Telomerase - metabolism
Telomeres
Templates, Genetic
thymidine
timidina
transferasas
transferase
transferases
Yeast
Yeasts
title Association of the Est1 protein with telomerase activity in yeast
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