Drosophila SPF45: A Bifunctional Protein with Roles in Both Splicing and DNA Repair

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Abstract
The sequence of the SPF45 protein is significantly conserved, yet functional studies have identified it as a splicing factor in animal cells and as a DNA-repair protein in plants. Using a combined genetic and biochemical approach to investigate this apparent functional discrepancy, we unify and validate both of these studies by demonstrating that the Drosophila melanogaster protein is bifunctional, with independent functions in DNA repair and splicing. We find that SPF45 associates with the U2 snRNP and that mutations that remove the C-terminal end of the protein disrupt this interaction. Although animals carrying this mutation are viable, they are nevertheless compromised in their ability to regulate Sex-lethal splicing, demonstrating that Sex-lethal is an important physiological target of SPF45. Furthermore, these mutant animals exhibit phenotypes diagnostic of difficulties in recovering from exogenously induced DNA damage. The conclusion that SPF45 functions in the DNA-repair pathway is strengthened by finding both genetic and physical interactions between SPF45 and RAD201, a previously uncharacterized member of the RecA/Rad51 protein family. Together with our finding that the fly SPF45 protein increases the survival rate of mutagen-treated bacteria lacking the RecG helicase, these studies provide the tantalizing suggestion that SPF45 has an ancient and evolutionarily conserved role in DNA repair. Assigning function to a protein relies on information about similar proteins in different species, and is based on the view that conservation of sequence generally parallels conservation of function. In this article, Chaouki and Salz focus on SPF45, a protein that, at first glance, appears to break this rule. Although the sequence of SPF45 is highly conserved, in animals cells SPF45 functions as a splicing factor, but in plant cells it functions as a DNA repair protein. This functional discrepancy is resolved here through the demonstration that, in D. melanogaster, SPF45 is a bifunctional protein with independent functions in DNA repair and splicing. Support for this conclusion includes the observation that mutant animals lacking SPF45 function display defects in both splicing and DNA repair. In addition, the authors show that SPF45 associates with two distinct groups of proteins; those that participate in RNA splicing and those that participate in DNA repair. The finding that the D. melanogaster protein is bifunctional suggests that the human protein may also have more than one function. This has important clinical implications because elevated SPF45 levels have been correlated with resistance to chemotherapy.

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