Association of the Est1 protein with telomerase activity in yeast

Steiner, B. R., Hidaka, K., Futcher, B. (1996) Association of the Est1 protein with telomerase activity in yeast. Proceedings of the National Academy of Sciences of the United States of America, 93 (7). pp. 2817-21. ISSN 0027-8424

URL: http://www.ncbi.nlm.nih.gov/pubmed/8610124

Abstract

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.

Item Type: Paper
Uncontrolled Keywords: Amino Acid Sequence Base Sequence Blotting, Northern Deoxyguanosine Fungal Proteins/isolation & purification/ metabolism Kinetics Molecular Sequence Data Mutagenesis Oligodeoxyribonucleotides/chemistry/metabolism RNA, Fungal/biosynthesis/isolation & purification/metabolism Research Support, Non-U.S. Gov't Saccharomyces cerevisiae/ enzymology/genetics Saccharomyces cerevisiae Proteins Substrate Specificity Telomerase/isolation & purification/ metabolism Templates, Genetic
Subjects: organism description > yeast > Saccharomyces
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > mutations > mutagenesis
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > telomerase
CSHL Authors:
Communities: CSHL labs > Futcher lab
Depositing User: Kathleen Darby
Date Deposited: 13 May 2014 13:48
Last Modified: 13 May 2014 13:48
Related URLs:
URI: http://repository.cshl.edu/id/eprint/30111

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