Tetrahymena proteins p80 and p95 are not core telomerase components.
Telomeres provide stability to eukaryotic chromosomes and consist of tandem DNA repeat sequences. Telomeric repeats are synthesized and maintained by a specialized reverse transcriptase, termed telomerase. Tetrahymena thermophila telomerase contains two essential components: Tetrahymena telomerase reverse transcriptase (tTERT), the catalytic protein component, and telomerase RNA that provides the template for telomere repeat synthesis. In addition to these two components, two proteins, p80 and p95, were previously found to copurify with telomerase activity and to interact with tTERT and telomerase RNA. To investigate the role of p80 and p95 in the telomerase enzyme, we tested the interaction of p80, p95, and tTERT in several different recombinant expression systems and in Tetrahymena extracts. Immunoprecipitation of recombinant proteins showed that although p80 and p95 associated with each other, they did not associate with tTERT. In in vitro transcription and translation lysates, tTERT was associated with telomerase activity, but p80 and p95 were not. p80 bound telomerase RNA as well as several other unrelated RNAs, suggesting p80 has a general affinity for RNA. Immunoprecipitations from Tetrahymena extracts also showed no evidence for an interaction between the core tTERT/telomerase RNA complex and the p80 and p95 proteins. These data suggest that p80 and p95 are not associated with the bulk of active telomerase in Tetrahymena.
Mason
D X
DX
Department of Molecular Biology and Genetics, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Autexier
C
C
Greider
C W
CW
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000502:physiology; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000737:chemistry; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000502:physiology; D011817:Rabbits; D019098:Telomerase / Q000737:chemistry*; D034501:Telomere-Binding Proteins
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D016601:RNA-Binding Proteins; D034501:Telomere-Binding Proteins; C416367:p95 protein, Tetrahymena; D019098:Telomerase; C498417:telomerase p80 protein, Tetrahymena