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J. Biol. Chem., Vol. 280, Issue 25, 23502-23515, June 24, 2005

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Comparative Functional Study of the Viral Telomerase RNA Based on Natural Mutations^*

Laetitia Fragnet, Emmanuel Kut, and Denis Rasschaert

From the Laboratoire Telomerase, Lymphome Viro-induit, Centre de Recherches INRA de Tours, UR-BASE 086, 37380 Nouzilly, France

Telomerase activity is present in most malignant tumors and provides a mechanism for the unlimited potential for division of neoplastic cells. We previously characterized the first identified viral telomerase RNA (vTR) encoded by the Marek's disease virus (MDV) (Fragnet, L., Blasco, M. A., Klapper, W., and Rasschaert, D. (2003) J. Virol. 77, 5985–5996). This avian herpesvirus induces T-lymphomas. We demonstrated that the vTR subunit of the oncogenic MDV-RB1B strain is functional and would be more efficient than its chicken counterpart, cTR, which is 88% homologous. We take advantage of the functionality of those natural mutant TRs to investigate the involvement of the mutations of vTR on its efficiency in a heterologous murine cell system and in a homologous in vitro system using the recombinant chicken telomerase reverse transcriptase. The P2 helix of the pseudoknot seems to be more stable in vTR than in cTR, and this may account for the higher activity of vTR than cTR. Moreover, the five adenines just upstream from the P3 helix of vTR may also play an important role in its efficiency. We also established that the substitution of a single nucleotide at the 3'-extremity of the H-box of the vaccine MDV-Rispens strain vTR resulted in a lack of its accumulation within the cell, especially in the nucleus, correlated with a decrease in telomerase activity.

Received for publication, February 1, 2005 , and in revised form, March 23, 2005.

^* This work was supported by grants from the "Ligue Nationale Contre le Cancer" and the "Région Centre." The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Laboratoire Telomerase, Lymphome Viro-induit, UR086, INRA, Centre de Recherches de Tours, 37380 Nouzilly, France. Tel.: 33-247-427-942; Fax: 33-247-427-774; E-mail: rasschae{at}tours.inra.fr.

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