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First published online 26 March 2008
doi: 10.1242/dev.020685

Development 135, 1659-1668 (2008)
Published by The Company of Biologists 2008
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Smt3 is required for Drosophila melanogaster metamorphosis

Ana Talamillo1,*, Jonatan Sánchez1,*, Rafael Cantera2,3, Coralia Pérez1, David Martín4, Eva Caminero5 and Rosa Barrio1,{dagger}

1 Functional Genomics Unit, CIC bioGUNE, Technology Park, Building 801-A, 48160 DERIO, Bizkaia, Spain.
2 Zoology Department, Stockholm University, 10691 Stockholm, Sweden.
3 Instituto de Investigaciones Biológicas Clemente Estable, Av. Italia, 3318 Montevideo, Uruguay.
4 Institut de Biologia Molecular de Barcelona, CSIC, J. Girona 18-26, 08034 Barcelona, Spain.
5 Centro de Biología Molecular Severo Ochoa, Universidad Autónoma de Madrid, 28049 Madrid, Spain.

{dagger} Author for correspondence (e-mail: rbarrio{at}cicbiogune.es)

Accepted 4 March 2008

Sumoylation, the covalent attachment of the small ubiquitin-related modifier SUMO to target proteins, regulates different cellular processes, although its role in the control of development remains unclear. We studied the role of sumoylation during Drosophila development by using RNAi to reduce smt3 mRNA levels in specific tissues. smt3 knockdown in the prothoracic gland, which controls key developmental processes through the synthesis and release of ecdysteroids, caused a 4-fold prolongation of larval life and completely blocked the transition from larval to pupal stages. The reduced ecdysteroid titer of smt3 knockdown compared with wild-type larvae explains this phenotype. In fact, after dietary administration of exogenous 20-hydroxyecdysone, knockdown larvae formed pupal cases. The phenotype is not due to massive cell death or degeneration of the prothoracic glands at the time when puparium formation should occur. Knockdown cells show alterations in expression levels and/or the subcellular localisation of enzymes and transcription factors involved in the regulation of ecdysteroid synthesis. In addition, they present reduced intracellular channels and a reduced content of lipid droplets and cholesterol, which could explain the deficit in steroidogenesis. In summary, our study indicates that Smt3 is required for the ecdysteroid synthesis pathway at the time of puparium formation.

Key words: Drosophila, Ecdysone, Metamorphosis, Ring gland, Smt3, Sumoylation






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