Open AccessCharacterization of human telomerase complex
Author(s) -
Shyam Ramakrishnan,
Harsh W. Sharma,
A. Darise Farris,
Kenneth M. Kaufman,
John B. Harley,
Kathleen Collins,
Ger J.M. Pruijn,
Walther J. van Venrooij,
Mitchell L. Martin,
Ramaswamy Narayanan
Publication year - 1997
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.94.19.10075
Subject(s) - telomerase , telomere , ribonucleoprotein , telomerase rna component , biology , rna , microbiology and biotechnology , protein subunit , polyclonal antibodies , dna , tetrahymena , immunoprecipitation , telomerase reverse transcriptase , antibody , genetics , gene
Telomerase, a ribonucleoprotein complex, adds hexameric repeats called “telomeres” to the growing ends of chromosomal DNA. Characterization of mammalian telomerase has been elusive because of its low level of expression. We describe a bioinformatics approach to enrich and characterize the human telomerase complex. Using local sequence homology search methods, we detected similarity of theTetrahymena p80 subunit of telomerase with the autoantigen Ro60. Antibodies to Ro60 immunoprecipitated the telomerase activity. Ro60 and p80 proteins were cross-recognizable by antibodies to either protein. Telomerase activity and the RNA component of telomerase complex were localized to a doublet in a native gel from the Ro60 antibody-precipitated material. The enriched material showed specific binding to a TTA GGG probein vitro in an RNA template-dependent manner. Polyclonal antibodies to the doublet also immunoprecipitated the telomerase activity. These results suggest an evolutionary conservation of the telomerase proteins.