The catalytic and the RNA subunits of human telomerase are required to immortalize equid primary fibroblastsReport as inadecuate




The catalytic and the RNA subunits of human telomerase are required to immortalize equid primary fibroblasts - Download this document for free, or read online. Document in PDF available to download.

Chromosoma

, Volume 121, Issue 5, pp 475–488

First Online: 14 July 2012Received: 07 April 2012Revised: 25 June 2012Accepted: 25 June 2012

Abstract

Many human primary somatic cells can be immortalized by inducing telomerase activity through the exogenous expression of the human telomerase catalytic subunit hTERT. This approach has been extended to the immortalization of cell lines from several mammals. Here, we show that hTERT expression is not sufficient to immortalize primary fibroblasts from three equid species, namely donkey, Burchelli’s zebra and Grevy’s zebra. In vitro analysis of a reconstituted telomerase composed by hTERT and an equid RNA component of telomerase TERC revealed a low activity of this enzyme compared to human telomerase, suggesting a low compatibility of equid and human telomerase subunits. This conclusion was also strengthened by comparison of human and equid TERC sequences, which revealed nucleotide differences in key regions for TERC and TERT interaction. We then succeeded in immortalizing equid fibroblasts by expressing hTERT and hTERC concomitantly. Expression of both human telomerase subunits led to telomerase activity and telomere elongation, indicating that human telomerase is compatible with the other equid telomerase subunits and proteins involved in telomere metabolism. The immortalization procedure described herein could be extended to primary cells from other mammals. The availability of immortal cells from endangered species could be particularly useful for obtaining new information on the organization and function of their genomes, which is relevant for their preservation.

Communicated by Jan Karlseder

Pamela Vidale tragically passed away on May 9, 2011. We miss her unique personality, intelligence, and passion for research.

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Author: Pamela Vidale - Elisa Magnani - Solomon G. Nergadze - Marco Santagostino - Gael Cristofari - Alexandra Smirnova - Chiara M

Source: https://link.springer.com/







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