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Published online before print August 30, 2004, 10.1261/rna.7122604
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RNA (2004), 10:1586-1594. Published by Cold Spring Harbor Laboratory Press. Copyright © 2004 RNA Society.

Trans-splicing and polyadenylation of let-7 microRNA primary transcripts

JOHN BRACHT1, SHAUN HUNTER1, RACHEL EACHUS, PHILLIP WEEKS and AMY E. PASQUINELLI

Department of Biology, University of California, San Diego, La Jolla, California 92093, USA

Reprint requests to: Amy E. Pasquinelli, Department of Biology, University of California, San Diego, La Jolla, CA 92093, USA; e-mail: apasquin{at}ucsd.edu. fax: (858) 822-3021.

Members of the microRNA (miRNA) class of 22-nucleotide RNAs regulate the expression of target genes that contain sequences of antisense complementarity. Maturation of miRNAs involves cleavage of longer primary transcripts, but little is yet understood about how miRNA genes are transcribed and enter the processing pathway. We find that relatively long, polyadenylated transcripts encoded by the Caenorhabditis elegans let-7 gene undergo trans-splicing to the spliced leader 1 (SL1) RNA. Deletions, including removal of the trans-splice site, upstream of mature let-7 sequence result in stable accumulation of primary transcripts and compromised production of mature let-7 RNA in vivo. Our data show that multiple steps of let-7 miRNA biogenesis can be uncoupled, allowing for complex regulation in the production of a functional miRNA. Finally, the observation that let-7 primary transcripts undergo splicing highlights the importance of identifying the sequence of endogenous pri-miRNA substrates recognized by the cellular processing machinery.

Keywords: microRNAs; let-7; trans-splicing; polyadenylation; miRNAs

Received June 25, 2004 ; accepted July 9, 2004.


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