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Molecular and Cellular Biology, August 2004, p. 6742-6750, Vol. 24, No. 15
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.15.6742-6750.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Dissection of a Natural RNA Silencing Process in the Drosophila melanogaster Germ Line

Alexei A. Aravin,1,{dagger} Mikhail S. Klenov,1 Vasilii V. Vagin,1 Frédéric Bantignies,2 Giacomo Cavalli,2 and Vladimir A. Gvozdev1*

Department of Animal Molecular Genetics, Institute of Molecular Genetics, Moscow 123182, Russia,1 Chromatin and Cell Biology Laboratory, Institute of Human Genetics, Centre National de la Recherche Scientifique, 34396 Montpellier, France2

Received 12 November 2003/ Returned for modification 31 December 2003/ Accepted 6 April 2004

To date, few natural cases of RNA-silencing-mediated regulation have been described. Here, we analyzed repression of testis-expressed Stellate genes by the homologous Suppressors of Stellate [Su(Ste)] repeats that produce sense and antisense short RNAs. The Stellate promoter is dispensable for suppression, but local disturbance of complementarity between the Stellate transcript and the Su(Ste) repeats impairs silencing. Using in situ RNA hybridization, we found temporal control of the expression and spatial distribution of sense and antisense Stellate and Su(Ste) transcripts in germinal cells. Antisense Su(Ste) transcripts accumulate in the nuclei of early spermatocytes before the appearance of sense transcripts. The sense and antisense transcripts are colocalized in the nuclei of mature spermatocytes, placing the initial step of silencing in the nucleus and suggesting formation of double-stranded RNA. Mutations in the aubergine and spindle-E genes, members of the Argonaute and RNA helicase gene families, respectively, impair silencing by eliminating the short Su(Ste) RNA, but have no effect on microRNA production. Thus, different small RNA-containing complexes operate in the male germ line.

* Corresponding author. Mailing address: Institute of Molecular Genetics of RAS, Kurchatov sq., 2, Moscow 123182, Russia. Phone: (7-095)196-0012. Fax: (7-095)196-0221. E-mail: gvozdev{at}img.ras.ru.

{dagger} Present address: Laboratory for RNA Molecular Biology, The Rockefeller University, New York, NY 10021.

Molecular and Cellular Biology, August 2004, p. 6742-6750, Vol. 24, No. 15
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.15.6742-6750.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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