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Molecular and Cellular Biology, February 2009, p. 861-870, Vol. 29, No. 3
0270-7306/09/$08.00+0     doi:10.1128/MCB.01524-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

A Drosophila pasha Mutant Distinguishes the Canonical MicroRNA and Mirtron Pathways{triangledown} ,{ddagger}

Raquel Martin,1,{dagger} Peter Smibert,1,{dagger} Abdullah Yalcin,2,§ David M. Tyler,1 Ulrich Schäfer,3 Thomas Tuschl,2 and Eric C. Lai1*

Sloan-Kettering Institute, Department of Developmental Biology, 1017 Rockefeller Research Laboratories, 1275 York Ave., Box 252, New York, New York 10065,1 Howard Hughes Medical Institute, Laboratory of RNA Molecular Biology, The Rockefeller University, 1230 York Ave., Box 186, New York, New York 10065-6399,2 Max-Planck-Institut fuer Biophysikalische Chemie, Abteilung Molekulare Entwicklungsbiologie, 37070 Goettingen, Germany3

Received 30 September 2008/ Returned for modification 30 October 2008/ Accepted 19 November 2008

Canonical primary microRNA (miRNA) transcripts and mirtrons are proposed to transit distinct nuclear pathways en route to generating mature ~22 nucleotide regulatory RNAs. We generated a null allele of Drosophila pasha, which encodes a double-stranded RNA-binding protein partner of the RNase III enzyme Drosha. Analysis of this mutant yielded stringent evidence that Pasha is essential for the biogenesis of canonical miRNAs but is dispensable for the processing and function of mirtron-derived regulatory RNAs. The pasha mutant also provided a unique tool to study the developmental requirements for Drosophila miRNAs. While pasha adult somatic clones are similar in many respects to those of dicer-1 clones, pasha mutant larvae revealed an unexpected requirement for the miRNA pathway in imaginal disc growth. These data suggest limitations to somatic clonal analysis of miRNA pathway components.

* Corresponding author. Mailing address: Sloan-Kettering Institute, Department of Developmental Biology, 1017 Rockefeller Research Laboratories, 1275 York Ave., Box 252, New York, NY 10065. Phone: (212) 639-5578. Fax: (212) 717-3604. E-mail: laie{at}mskcc.org

{triangledown} Published ahead of print on 1 December 2008.

{ddagger} Supplemental material for this article may be found at http://mcb.asm.org/.

{dagger} R.M. and P.S. contributed equally to this study.

§ Present address: Department of Genetics and Complex Diseases, Harvard School of Public Health, 655 Huntington Ave., SPH I-210, Boston, MA 02115.

Molecular and Cellular Biology, February 2009, p. 861-870, Vol. 29, No. 3
0270-7306/09/$08.00+0     doi:10.1128/MCB.01524-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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