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Nucleic Acids Research 2005 33(2):464-477; doi:10.1093/nar/gki171
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Published online 19 January 2005

© 2005, the authors Nucleic Acids Research, Vol. 33 No. 2 © Oxford University Press 2005; all rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use permissions, please contact journals.permissions{at}oupjournals.org.

Article

Conserved and specific functions of mammalian ssu72

Benoit St-Pierre1, Xudong Liu1, Lan-Chau T. Kha1,4, Xudong Zhu1, Owen Ryan5, Zhe Jiang1 and Eldad Zacksenhaus1,2,3,4,*

1 Division of Cell and Molecular Biology, Toronto General Research Institute, University Health Network 67 College Street, Room 407, Toronto, Ontario, Canada M5G 2m1 2 Department of Medicine, University of Toronto Toronto, Ontario, Canada M5G 2M1 3 Department of Laboratory Medicine and Pathobiology, University of Toronto Toronto, Ontario, Canada M5G 2M1 4 Department of Medical Biophysics, University of Toronto Toronto, Ontario, Canada M5G 2M1 5 Banting and Best Department of Medical Research Toronto, Ontario, Canada M5G 1L6

*To whom correspondence should be addressed. Tel: +1 416 340 4800 ext. 5106; Fax: +1 416 340 3453; Email: eldad.zacksenhaus{at}utoronto.ca

Received October 27, 2004. Revised December 16, 2004. Accepted December 16, 2004.

We describe the cloning and characterization of a human homolog of the yeast transcription/RNA-processing factor Ssu72, following a yeast two-hybrid screen for pRb-binding factors in the prostate gland. Interaction between hSsu72 and pRb was observed in transfected mammalian cells and involved multiple domains in pRb; however, so far, mutual effects of these two factors could not be demonstrated. Like the yeast counterpart, mammalian Ssu72 associates with TFIIB and the yeast cleavage/polyadenylation factor Pta1, and exhibits intrinsic phosphatase activity. Mammals contain a single ssu72 gene and a few pseudogenes. During mouse embryogenesis, ssu72 was highly expressed in the nervous system and intestine; high expression in the nervous system persisted in adult mice and was also readily observed in multiple human tumor cell lines. Both endogenous and ectopically expressed mammalian Ssu72 proteins resided primarily in the cytoplasm and only partly in the nucleus. Interestingly, fusion to a strong nuclear localization signal conferred nuclear localization only in a fraction of transfected cells, suggesting active tethering in the cytoplasm. Suppression of ssu72 expression in mammalian cells by siRNA did not reduce proliferation/survival, and its over-expression did not affect transcription of candidate genes in transient reporter assays. Despite high conservation, hssu72 was unable to rescue an ssu72 lethal mutation in yeast. Together, our results highlight conserved and mammalian specific characteristics of mammalian ssu72.

The authors wish it to be known that, in their opinion, the first four authors should be regarded as joint First Authors


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