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MBC in Press, published online ahead of print February 28, 2002
Mol. Biol. Cell 10.1091/mbc.01-09-0436

A more recent version of this article appeared on May 1, 2002
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Submitted on August 31, 2001
Revised on December 31, 2001
Accepted on February 11, 2002

Evidence for a Post-Transcriptional Role of a TFIIIC{alpha}-like protein in Chironomus tentans

Nafiseh Sabri1, Ann-Kristin Östlund Farrants2, Ulf Hellman3, and Neus Visa1*

1 Department of Molecular Biology and Functional Genomics, Stockholm University, SE-10691 Stockholm, Sweden
2 Department of Zoological Cell Biology, The Wenner-Gren Institute, Stockholm University, SE-10691 Stockholm, Sweden
3 Ludwig Institute for Cancer Research, Box 595, SE-751 24 Uppsala, Sweden

* Corresponding author. E-mail address: Neus.Visa{at}molbio.su.se.

We have cloned and sequenced a cDNA that encodes for a nuclear protein of 238 kDa in the dipteran Chironomus tentans. This protein, that we call p2D10, is structurally similar to the {alpha} subunit of the general transcription factor TFIIIC. Using immuno-electron microscopy we have shown that a fraction of p2D10 is located at sites of transcription, which is consistent with a possible role of this protein in transcription initiation. We have also found that a large fraction of p2D10 is located in the nucleoplasm and in the nuclear pore complexes. Using gel filtration chromatography and co-immunoprecipitation methods, we have identified and characterized two p2D10-containing complexes that differ in molecular mass and composition. The heavy p2D10-containing complex contains at least one other component of the TFIIIC complex, TFIIIC-{epsilon}. Based on its molecular mass and composition, the heavy p2D10-containing complex may be the Pol III holoenzyme. The light p2D10-containing complex contains RNA together with at least two proteins that are thought to be involved in mRNA trafficking, RAE1 and hrp65. The observations reported here suggest that this new TFIIIC-{alpha}-like protein is involved in post-transcriptional steps of pre-mRNA metabolism in Chironomus tentans.




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