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Molecular and Cellular Biology, May 2004, p. 4229-4240, Vol. 24, No. 10
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.10.4229-4240.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

The Splicing Factor U2AF Small Subunit Is Functionally Conserved between Fission Yeast and Humans

Christopher J. Webb and Jo Ann Wise*

Department of Molecular Biology and Microbiology, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106-4960

Received 20 November 2003/ Returned for modification 17 December 2003/ Accepted 25 February 2004

The small subunit of U2AF, which functions in 3' splice site recognition, is more highly conserved than its heterodimeric partner yet is less thoroughly investigated. Remarkably, we find that the small subunit of Schizosaccharomyces pombe U2AF (U2AFSM) can be replaced in vivo by its human counterpart, demonstrating that the conservation extends to function. Precursor mRNAs accumulate in S. pombe following U2AFSM depletion in a time frame consistent with a role in splicing. A comprehensive mutational analysis reveals that all three conserved domains are required for viability. Notably, however, a tryptophan in the pseudo-RNA recognition motif implicated in a key contact with the large subunit by crystallographic data is dispensable whereas amino acids implicated in RNA recognition are critical. Mutagenesis of the two zinc-binding domains demonstrates that they are neither equivalent nor redundant. Finally, two- and three-hybrid analyses indicate that mutations with effects on large-subunit interactions are rare whereas virtually all alleles tested diminished RNA binding by the heterodimer. In addition to demonstrating extraordinary conservation of U2AF small-subunit function, these results provide new insights into the roles of individual domains and residues.

* Corresponding author. Mailing address: Department of Molecular Biology and Microbiology, School of Medicine, Case Western Reserve University, 10900 Euclid Ave., Cleveland, OH 44106-4960. Phone: (216) 368-1876. Fax: (216) 368-3055. E-mail: jaw17{at}case.edu.

Molecular and Cellular Biology, May 2004, p. 4229-4240, Vol. 24, No. 10
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.10.4229-4240.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Sridharan, V., Singh, R. (2007). A Conditional Role of U2AF in Splicing of Introns with Unconventional Polypyrimidine Tracts. Mol. Cell. Biol. 27: 7334-7344 [Abstract] [Full Text]  
  • Haraguchi, N., Andoh, T., Frendewey, D., Tani, T. (2007). Mutations in the SF1-U2AF59-U2AF23 Complex Cause Exon Skipping in Schizosaccharomyces pombe. J. Biol. Chem. 282: 2221-2228 [Abstract] [Full Text]  
  • Wang, B.-B., Brendel, V. (2006). Molecular Characterization and Phylogeny of U2AF35 Homologs in Plants. Plant Physiol. 140: 624-636 [Abstract] [Full Text]  
  • Webb, C. J., Lakhe-Reddy, S., Romfo, C. M., Wise, J. A. (2005). Analysis of Mutant Phenotypes and Splicing Defects Demonstrates Functional Collaboration between the Large and Small Subunits of the Essential Splicing Factor U2AF In Vivo. Mol. Biol. Cell 16: 584-596 [Abstract] [Full Text]  
  • Webb, C. J., Romfo, C. M., van Heeckeren, W. J., Wise, J. A. (2005). Exonic splicing enhancers in fission yeast: functional conservation demonstrates an early evolutionary origin. Genes Dev. 19: 242-254 [Abstract] [Full Text]  
  • Yashiroda, H., Tanaka, K. (2004). Hub1 is an essential ubiquitin-like protein without functioning as a typical modifier in fission yeast. GENES CELLS 9: 1189-1197 [Abstract] [Full Text]  


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