Characterization of human telomerase complex.
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 the Tetrahymena 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 probe in 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.
Ramakrishnan
S
S
Department of Oncology, Hoffmann-La Roche, 340 Kingsland Street, Nutley, NJ 07110, USA.
Sharma
H W
HW
Farris
A D
AD
Kaufman
K M
KM
Harley
J B
JB
Collins
K
K
Pruijn
G J
GJ
van Venrooij
W J
WJ
Martin
M L
ML
Narayanan
R
R
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D001324:Autoantigens / Q000737:chemistry / Q000276:immunology; D001666:Binding Sites, Antibody; D002461:Cell Line, Transformed; D003429:Cross Reactions; D006801:Humans; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D019098:Telomerase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D013768:Tetrahymena / Q000201:enzymology; D014407:Tumor Cells, Cultured
D000906:Antibodies; D001324:Autoantigens; D019098:Telomerase