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First published online 13 June 2006
doi: 10.1242/jcs.03001

Journal of Cell Science 119, 2797-2806 (2006)
Published by The Company of Biologists 2006
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Research Article

Immunodetection of human telomerase reverse-transcriptase (hTERT) re-appraised: nucleolin and telomerase cross paths

Ying-Li Wu1,2, Charles Dudognon1, Eric Nguyen1, Josette Hillion1, Frédéric Pendino1, Ilona Tarkanyi3, Janos Aradi3, Michel Lanotte1, Jian-Hua Tong4, Guo-Qiang Chen2 and Evelyne Ségal-Bendirdjian1,*

1 INSERM U685, Hôpital Saint-Louis, Institut d'Hématologie, 1 avenue Claude Vellefaux, 75010 Paris, France
2 Department of Pathophysiology Key Laboratory of Cell Differentiation and Apoptosis of Ministry of Education, Shanghai Jiao-Tong University School of Medicine, Shanghai 200025, P. R. China
3 Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, Nagyerdei krt. 98, 4012 Debrecen, Hungary
4 Shanghai Institute of Hematology, Ruijin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai 200025, P. R. China

* Author for correspondence (e-mail: segal{at}stlouis.inserm.fr)

Accepted 29 March 2006

The involvement of telomerase in cellular immortalization and senescence has often been assessed by means of telomerase expression at the RNA level and quantification of telomerase activity by the telomeric repeat amplification protocol assay. However, these methods either neglected the existence of various telomerase splice variants, or ignored the nonconventional functions of telomerase independent of its ability to elongate and maintain telomere length. Immunodetection of telomerase is now being recognized as a necessary approach to precisely elucidate its roles in oncogenesis and senescence. A few antibodies directed against the catalytic subunit of the human telomerase (hTERT) are currently used but their specificity is not always demonstrated. A survey of the literature showed inconsistencies and led us to comparatively re-evaluate the most frequently used antibodies. Surprisingly, mass spectrometry, two-dimensional gel analysis and immunofluorescent experiments revealed that the most frequently used hTERT immunoprobe, a mouse monoclonal antibody that was claimed to be directed against an hTERT protein epitope, in fact recognizes nucleolin rather than telomerase. Our findings have interesting implications regarding the biology of nucleolin and telomerase in the context of pathophysiological investigations recently carried out.

Key words: Telomerase, Nucleolin, Immunodetection






© The Company of Biologists Ltd 2006