Cloning and intracellular localization of the U2 small nuclear ribonucleoprotein auxiliary factor small subunit.

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Proceedings of the National Academy of Sciences of the United States of America, ISSN: 0027-8424, Vol: 89, Issue: 18, Page: 8769-73

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Zhang, Meng; Zamore, Phillip D.; Carmo-Fonseca, Maria; Lamond, Angus I.; Green, Michael R.
Proceedings of the National Academy of Sciences
Multidisciplinary; Life Sciences; Medicine and Health Sciences
article description
U2 small nuclear ribonucleoprotein auxiliary factor (U2AF), an essential mammalian splicing factor, is composed of two subunits: a 65-kDa protein (U2AF65), which binds the pre-mRNA polypyrimidine tract and is required for in vitro splicing, and an associated 35-kDa protein (U2AF35). Here we report the isolation of a cDNA encoding U2AF35. U2AF35 contains sequence motifs found in several mammalian pre-mRNA splicing factors. We show directly that U2AF65 and U2AF35 interact with each other and delineate the regions of both proteins that mediate this interaction. Using anti-peptide antibodies against U2AF35, we show that the protein has the intracellular distribution characteristic of U2AF65. Both U2AF65 and U2AF35 are concentrated in a small number of nuclear foci corresponding to coiled bodies, subnuclear organelles first identified by light microscopy in 1903.