Protein composition of catalytically active human telomerase from immortal cells.
Telomerase is a ribonucleoprotein enzyme complex that adds 5'-TTAGGG-3' repeats onto the ends of human chromosomes, providing a telomere maintenance mechanism for approximately 90% of human cancers. We have purified human telomerase approximately 10(8)-fold, with the final elution dependent on the enzyme's ability to catalyze nucleotide addition onto a DNA oligonucleotide of telomeric sequence, thereby providing specificity for catalytically active telomerase. Mass spectrometric sequencing of the protein components and molecular size determination indicated an enzyme composition of two molecules each of telomerase reverse transcriptase, telomerase RNA, and dyskerin.
Cohen
Scott B
SB
Cancer Research Unit, Children's Medical Research Institute, 214 Hawkesbury Road, Westmead NSW 2145, Australia.
Graham
Mark E
ME
Lovrecz
George O
GO
Bache
Nicolai
N
Robinson
Phillip J
PJ
Reddel
Roger R
RR
D000595:Amino Acid Sequence; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002460:Cell Line; D045744:Cell Line, Tumor; D002499:Centrifugation, Density Gradient; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000737:chemistry; D009687:Nuclear Proteins / Q000737:chemistry* / Q000302:isolation & purification; D012313:RNA / Q000737:chemistry* / Q000302:isolation & purification; D053719:Tandem Mass Spectrometry; D019098:Telomerase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
D018797:Cell Cycle Proteins; C112148:DKC1 protein, human; D009097:Multienzyme Complexes; D009687:Nuclear Proteins; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase