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MBC in Press, published online ahead of print December 7, 2002
Mol. Biol. Cell 10.1091/mbc.E02-09-0601

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Submitted on September 19, 2002
Revised on October 30, 2002
Accepted on November 22, 2002

Subnuclear localization and Cajal body targeting of transcription elongation factor TFIIS in amphibian oocytes

Abigail J. Smith1, Yan Ling1, and Garry T. Morgan1*

1 Institute of Genetics, University of Nottingham, Nottingham NG7 2UH, UK

* Corresponding author. E-mail address: garry.morgan{at}nottingham.ac.uk.

We have examined the localization and targeting of the RNA polymerase II (pol II) transcription elongation factor TFIIS in amphibian oocyte nuclei by immunofluorescence. Using a novel antibody against Xenopus TFIIS the major sites of immunostaining were found to be Cajal bodies, nuclear organelles that also contain pol II. Small granular structures attached to lampbrush chromosomes were also specifically stained but the transcriptionally-active loops were not. Similar localization patterns were found for the newly-synthesised myc-tagged TFIIS produced after injection of synthetic transcripts into the cytoplasm. The basis of the rapid and preferential targeting of TFIIS to Cajal bodies was investigated by examining the effects of deletion and site-specific mutations. Multiple regions of TFIIS contributed to efficient targeting including the domain required for its binding to pol II. The localisation of TFIIS in Cajal bodies, and in particular the apparent involvement of pol II binding in achieving it, offer further support for a model in which Cajal bodies function in the pre-assembly of the transcriptional machinery. Although our findings are therefore consistent with TFIIS playing a role in early events of the transcription cycle they also suggest that this elongation factor is not generally required during transcription in oocytes.




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