Two inactive fragments of the integral RNA cooperate to assemble active telomerase with the human protein catalytic subunit (hTERT) in vitro

Valerie M. Tesmer, Lance P. Ford, Shawn E. Holt, Bryan C. Frank, Xiaoming Yi, Dara L. Aisner, Michel M Ouellette, Jerry W. Shay, Woodring E. Wright

Research output: Contribution to journalArticle

104 Citations (Scopus)

Abstract

We have mapped the 5' and 3' boundaries of the region of the human telomerase RNA (hTR) that is required to produce activity with the human protein catalytic subunit (hTERT) by using in vitro assembly systems derived from rabbit reticulocyte lysates and human cell extracts. The region spanning nucleotides +33 to +325 of the 451-base hTR is the minimal sequence required to produce levels of telomerase activity that are comparable with that made with full-length hTR. Our results suggest that the sequence approximately 270 bases downstream of the template is required for efficient assembly of active telomerase in vitro; this sequence encompasses a substantially larger portion of the 3' end of hTR than previously thought necessary. In addition, we identified two fragments of hTR (nucleotides +33 to +147 and +164 to +325) that cannot produce telomerase activity when combined separately with hTERT but can function together to assemble active telomerase. These results suggest that the minimal sequence of hTR can be divided into two sections, both of which are required for de novo assembly of active telomerase in vitro.

Original languageEnglish (US)
Pages (from-to)6207-6216
Number of pages10
JournalMolecular and cellular biology
Volume19
Issue number9
DOIs
StatePublished - Jan 1 1999

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Telomerase
Protein Subunits
Catalytic Domain
RNA
Nucleotides
Reticulocytes
In Vitro Techniques
Cell Extracts
Human Activities
telomerase RNA
Rabbits

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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Two inactive fragments of the integral RNA cooperate to assemble active telomerase with the human protein catalytic subunit (hTERT) in vitro. / Tesmer, Valerie M.; Ford, Lance P.; Holt, Shawn E.; Frank, Bryan C.; Yi, Xiaoming; Aisner, Dara L.; Ouellette, Michel M; Shay, Jerry W.; Wright, Woodring E.

In: Molecular and cellular biology, Vol. 19, No. 9, 01.01.1999, p. 6207-6216.

Research output: Contribution to journalArticle

Tesmer, Valerie M. ; Ford, Lance P. ; Holt, Shawn E. ; Frank, Bryan C. ; Yi, Xiaoming ; Aisner, Dara L. ; Ouellette, Michel M ; Shay, Jerry W. ; Wright, Woodring E. / Two inactive fragments of the integral RNA cooperate to assemble active telomerase with the human protein catalytic subunit (hTERT) in vitro. In: Molecular and cellular biology. 1999 ; Vol. 19, No. 9. pp. 6207-6216.
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