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| Subject Categories: Membranes & Transport | RNA |
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| The EMBO Journal
(2008) 27, 748–757, doi:10.1038/emboj.2008.21 Published online 14 February 2008 |
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| TLC1 RNA nucleo-cytoplasmic trafficking links telomerase biogenesis to its recruitment to telomeres |
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| Franck Gallardo1, 3, Catherine Olivier1, 3, Alain T Dandjinou2, Raymund J Wellinger2 and Pascal Chartrand1 |
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1 Département de Biochimie, Université de Montréal, Montréal, Quebec, Canada 2 Département de Microbiologie et Infectiologie, Université de Sherbrooke, Sherbrooke, Quebec, Canada To whom correspondence should be addressed Pascal Chartrand, Department of Biochemistry, Université de Montréal, 2900 Edouard-Montpetit, Montréal, Quebec, Canada H3C 3J7. Tel.: +1 514 343 5684; Fax: +1 514 343 2210; E-mail: p.chartrand@umontreal.ca 3 These authors contributed equally to this work Received 24 October 2007; Accepted 21 January 2008; Published online 14 February 2008. |
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| Abstract |
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| The yeast telomerase holoenzyme, which adds telomeric repeats at the chromosome ends, is composed of the TLC1 RNA and the associated proteins Est1, Est2 and Est3. To study the biogenesis of telomerase in endogenous conditions, we performed fluorescent in situ hybridization on the native TLC1 RNA. We found that the telomerase RNA colocalizes with telomeres in G1- to S-phase cells. Strains lacking any one of the Est proteins accumulate TLC1 RNA in their cytoplasm, indicating that a critical stage of telomerase biogenesis could take place outside of the nucleus. We were able to demonstrate that endogenous TLC1 RNA shuttles between the nucleus and the cytoplasm, in association with the Crm1p exportin and the nuclear importins Mtr10p–Kap122p. Furthermore, nuclear retention of the TLC1 RNA is impaired in the absence of yKu70p, Tel1p or the MRX complex, which recruit telomerase to telomeres. Altogether, our results reveal that the nucleo-cytoplasmic trafficking of the TLC1 RNA is an important step in telomere homeostasis, and link telomerase biogenesis to its recruitment to telomeres. |
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| Keywords: Crm1p, FISH, nucleo-cytoplasmic shuttling, telomerase, TLC1 RNA |
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