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Molecular and Cellular Biology, February 2001, p. 1111-1120, Vol. 21, No. 4
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.4.1111-1120.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Intercistronic Region Required for Polycistronic Pre-mRNA Processing in Caenorhabditis elegans

Tao Huang,dagger Scott Kuersten,Dagger Atul M. Deshpande, John Spieth,§ Margaret MacMorris, and Thomas Blumenthal*

Department of Biochemistry and Molecular Genetics, University of Colorado Health Sciences Center, Denver, Colorado 80262

Received 17 October 2000/Returned for modification 3 November 2000/Accepted 8 November 2000

In Caenorhabditis elegans, polycistronic pre-mRNAs are processed by cleavage and polyadenylation at the 3' ends of the upstream genes and trans splicing, generally to the specialized spliced leader SL2, at the 5' ends of the downstream genes. Previous studies have indicated a relationship between these two events in the processing of a heat shock-induced gpd-2-gpd-3 polycistronic pre-mRNA. Here, we report mutational analysis of the intercistronic region of this operon by linker scan analysis. Surprisingly, no sequences downstream of the 3' end were important for 3'-end formation. In contrast, a U-rich (Ur) element located 29 bp downstream of the site of 3'-end formation was shown to be important for downstream mRNA biosynthesis. This ~20-bp element is sufficient for SL2 trans splicing and mRNA accumulation when transplanted to a heterologous context. Furthermore, when the downstream gene was replaced by a gene from another organism, no loss of trans-splicing specificity was observed, suggesting that the Ur element may be the primary signal required for downstream mRNA processing.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Genetics, University of Colorado Health Sciences Center, Box B-121, 4200 E. 9th Ave., Denver CO 80262. Phone: (303) 315-8181. Fax: (303) 315-8215. E-mail: tom.blumenthal{at}uchsc.edu.

dagger Present address: Department of Molecular Biology, Princeton University, Princeton, NJ 08544.

Dagger Present address: Gene Expression Program, EMBL, D-69117 Heidelberg, Germany.

§ Present address: Genome Sequencing Center, Washington University School of Medicine, St. Louis, MO 63108.

Molecular and Cellular Biology, February 2001, p. 1111-1120, Vol. 21, No. 4
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.4.1111-1120.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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