Gel shift and UV cross-linking analysis of Tetrahymena telomerase

Harrington, L., Hull, C., Crittenden, J., Greider, C. (April 1995) Gel shift and UV cross-linking analysis of Tetrahymena telomerase. J Biol Chem, 270 (15). pp. 8893-901. ISSN 0021-9258 (Print)

URL: http://www.ncbi.nlm.nih.gov/pubmed/7721797
DOI: 10.1074/jbc.270.15.8893

Abstract

Telomerase is an unusual ribonucleoprotein that synthesizes new telomeres onto chromosome ends. The enzyme has been most extensively characterized in ciliates, where the RNA component has been cloned from several species, and its elongation properties have been characterized in detail. To understand the substrate specificity and protein composition of telomerase, we have used gel shift and UV cross-linking to characterize the enzyme from the ciliate Tetrahymena thermophila. In a mobility shift assay, a complex was identified that contained telomerase RNA, co-purified with telomerase activity, and was sensitive to nuclease treatment. The mobility shift complexes specifically formed using several different single-stranded, telomeric sequences but not non-telomeric primers. These results suggest that the specificity of telomerase for G-rich primer sequences occurs at least in part at the level of primer binding. UV cross-linking analysis identified a 100-kDa cross-linked protein that may be a telomerase component.

Item Type: Paper
Uncontrolled Keywords: Animals Base Sequence Chromatography, Gel Chromatography, Ion Exchange DNA/metabolism DNA Nucleotidylexotransferase/ chemistry/isolation & purification/metabolism/radiation effects DNA Primers Electrophoresis, Agar Gel Electrophoresis, Polyacrylamide Gel Molecular Sequence Data Protein Binding Substrate Specificity Tetrahymena thermophila/ enzymology Ultraviolet Rays
Subjects: organism description > animal
biotechnology > chromatography
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > DNA binding protein
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > telomerase
CSHL Authors:
Communities: CSHL labs
Depositing User: Jessica Koos
Date: 14 April 1995
Date Deposited: 11 Aug 2014 16:11
Last Modified: 11 Aug 2014 16:11
Related URLs:
URI: http://repository.cshl.edu/id/eprint/30600

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