Author(s): Custódio, Noélia ; Antoniou, Michael ; Carmo-Fonseca, Maria
Date: 2006
Persistent ID: http://hdl.handle.net/10451/47751
Origin: Repositório da Universidade de Lisboa
Subject(s): RNA polymerase II; Nuclear export; CRM1; Nucleo-cytoplasmic shuttling
© 2006 Elsevier Inc. All rights reserved.
Eukaryotic RNA polymerase II is a complex enzyme composed of 12 distinct subunits that is present in cells in low abundance. Transcription of mRNA by RNA polymerase II involves a phosphorylation/dephosphorylation cycle of the carboxyl-terminal domain (CTD) of the enzyme's largest subunit. We have generated stable murine cell lines expressing an alpha-amanitin-resistant form of the largest subunit of RNA polymerase II (RNA Pol II LS). These cells maintained transcriptional activity in the presence of alpha-amanitin, indicating that the exogenous protein was functional. We observed that over-expressed RNA Pol II LS was predominantly hypophosphorylated, soluble and accumulated in the cytoplasm in a CRM1-dependent manner. Our results further showed that the transcriptionally active form of RNA Pol II LS containing phosphoserine in position 2 of the CTD repeats was restricted to the nucleus and its levels remained remarkably constant. We propose that nucleo-cytoplasmic shuttling of RNA Pol II LS may provide a mechanism to control the pool of RNA polymerase subunits that is accessible for assembly of a functional enzyme in the nucleus.
This work was supported by Fundação para a Ciência e Tecnologia, Portugal, and by the European Science Foundation (EuroDYNA Programme)
