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Eukaryotic Cell, June 2006, p. 924-934, Vol. 5, No. 6
1535-9778/06/$08.00+0     doi:10.1128/EC.00065-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Identification of the Major Spliceosomal RNAs in Dictyostelium discoideum Reveals Developmentally Regulated U2 Variants and Polyadenylated snRNAs

Andrea Hinas,^1, Pontus Larsson,^2, Lotta Avesson,¹ Leif A. Kirsebom,² Anders Virtanen,² and Fredrik Söderbom^1*

Department of Molecular Biology, Biomedical Center, Swedish University of Agricultural Sciences, Box 590, SE-75124 Uppsala, Sweden,¹ Department of Cell and Molecular Biology, Biomedical Center, Uppsala University, Box 596, SE-75124 Uppsala, Sweden²

Received 3 March 2006/ Accepted 3 April 2006

Most eukaryotic mRNAs depend upon precise removal of introns by the spliceosome, a complex of RNAs and proteins. Splicing of pre-mRNA is known to take place in Dictyostelium discoideum, and we previously isolated the U2 spliceosomal RNA experimentally. In this study, we identified the remaining major spliceosomal RNAs in Dictyostelium by a bioinformatical approach. Expression was verified from 17 small nuclear RNA (snRNA) genes. All these genes are preceded by a putative noncoding RNA gene promoter. Immunoprecipitation showed that snRNAs U1, U2, U4, and U5, but not U6, carry the conserved trimethylated 5' cap structure. A number of divergent U2 species are expressed in Dictyostelium. These RNAs carry the U2 RNA hallmark sequence and structure motifs but have an additional predicted stem-loop structure at the 5' end. Surprisingly, and in contrast to the other spliceosomal RNAs in this study, the new U2 variants were enriched in the cytoplasm and were developmentally regulated. Furthermore, all of the snRNAs could also be detected as polyadenylated species, and polyadenylated U1 RNA was demonstrated to be located in the cytoplasm.

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* Corresponding author. Mailing address: Department of Molecular Biology, Biomedical Center, Swedish University of Agricultural Sciences, Box 590, SE-75124 Uppsala, Sweden. Phone: 46 18 471 4901. Fax: 46 18 536971. E-mail: fredde{at}xray.bmc.uu.se.

Supplemental material for this article may be found at http://ec.asm.org/.

These authors contributed equally to this work.

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Eukaryotic Cell, June 2006, p. 924-934, Vol. 5, No. 6
1535-9778/06/$08.00+0     doi:10.1128/EC.00065-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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