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Science 30 January 2004:
Vol. 303. no. 5658, pp. 669 - 672
DOI: 10.1126/science.1092653
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Reports

Heterochromatic Silencing and HP1 Localization in Drosophila Are Dependent on the RNAi Machinery

Manika Pal-Bhadra,1,3* Boris A. Leibovitch,2* Sumit G. Gandhi,3 Madhusudana Rao,3 Utpal Bhadra,1,3{dagger} James A. Birchler,1{dagger} Sarah C. R. Elgin2{dagger}

Genes normally resident in euchromatic domains are silenced when packaged into heterochromatin, as exemplified in Drosophila melanogaster by position effect variegation (PEV). Loss-of-function mutations resulting in suppression of PEV have identified critical components of heterochromatin, including proteins HP1, HP2, and histone H3 lysine 9 methyltransferase. Here, we demonstrate that this silencing is dependent on the RNA interference machinery, using tandem mini-white arrays and white transgenes in heterochromatin to show loss of silencing as a result of mutations in piwi, aubergine, or spindle-E (homeless), which encode RNAi components. These mutations result in reduction of H3 Lys9 methylation and delocalization of HP1 and HP2, most dramatically in spindle-E mutants.

1 Division of Biological Sciences, 117 Tucker Hall, University of Missouri, Columbia, MO 65211, USA.
2 Department of Biology, Washington University, St. Louis, MO 63130, USA.
3 Functional Genomics and Gene Silencing Group, Centre for Cellular and Molecular Biology, Hyderabad 500007, India.



* These authors contributed equally to this work.

{dagger} To whom correspondence should be addressed. The three laboratories contributed equally. E-mail: selgin{at}biology.wustl.edu, birchlerj{at}missouri.edu, utpal{at}ccmb.res.in.

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K.A. HAYNES, B.A. LEIBOVITCH, S.H. RANGWALA, C. CRAIG, and S.C.R. ELGIN (2004)
Cold Spring Harb Symp Quant Biol 69, 267-272
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Poetry of b1 Paramutation: cis- and trans-Chromatin Communication.
V.L. CHANDLER (2004)
Cold Spring Harb Symp Quant Biol 69, 355-362
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Regulation of Caenorhabditis elegans RNA Interference by the daf-2 Insulin Stress and Longevity Signaling Pathway.
D. WANG and G. RUVKUN (2004)
Cold Spring Harb Symp Quant Biol 69, 429-432
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Interrelationship of RNA Interference and Transcriptional Gene Silencing in Drosophila.
M. PAL-BHADRA, U. BHADRA, and J.A. BIRCHLER (2004)
Cold Spring Harb Symp Quant Biol 69, 433-438
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Science. ISSN 0036-8075 (print), 1095-9203 (online)