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MBC in Press, published online ahead of print July 21, 2004
Mol. Biol. Cell 10.1091/mbc.E04-02-0138

A more recent version of this article appeared on October 1, 2004
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Submitted on February 19, 2004
Revised on June 9, 2004
Accepted on June 30, 2004

Cell-specific Nucleolar Localisation of TBP-related Factor 2

Philippe Kieffer-Kwon, Igor Martianov*, and Irwin Davidson{dagger}

Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, 67404 Illkirch Cédex., France

Monitoring Editor: Keith Yamamoto

TATA-binding protein (TBP)-related factor 2 (TRF2) is one of four closely related RNA polymerase II transcription factors. We compared the intracellular localizations of TBP and TRF2 during the cell cycle and mitosis in HeLa cells. We show that during interphase, endogenous or exogenously expressed TRF2 is located almost exclusively in the nucleolus in HeLa or Cos cells. TRF2 localization is not affected by stress or mitotic stimuli, but TRF2 is rapidly released from the nucleolus upon inhibition of pol I transcription or treatment by RNase. These results suggest that localization of HeLa TRF2 requires a nucleolar-associated RNA species. In contrast, in 3T3 fibroblast cells, exogenously expressed TRF2 localizes to the nucleoplasm. Constitutive expression of ectopic TRF2 in 3T3 cells leads to a prolonged S phase of the cell cycle and reduced proliferation. Together with previous data, our results highlight the cell-specific localization and functions of TRF2. Furthermore, we show that during cell division, HeLa TRF2 and TBP are localized in the mitotic cytoplasm and TRF2 relocalises into the nascent nucleoli immediately after mitosis, whereas TBP reassociates with the chromatin. Although partially contradictory results have been reported, our data consistent with a model where only small proportion of the cellular TBP remains associated with specific promoter loci during mitosis.

*Present address: Sir William Dunn School of Pathology, South Parks Rd., Oxford, United Kingdom

{dagger}Corresponding author. E-mail: irwin{at}titus.u-strasbg.fr




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