Drosophila genome-wide regulatorsRNAi screen identifies multiplesregulators of HIF dependent transcription in hypoxia

JOEL PEREZ-PERRI; ANDRÉS DEKANTY; NURIA M. ROMERO; AGUSTINA BERTOLIN

Colorado, Estados Unidos

Simposio; Hypoxia: Molecular mechanism of oxygen sensing and response pathways; 2010

Keystone Symposia on molecular and cellular Biology

Drosophila genome-wide RNAi screen identifies multiple regulators of HIF dependent transcription in hypoxia   Joel I. Perez-Perri1, Andrés Dekanty1, Nuria Romero1, Agostina Bertolin1, Pablo Wappner1.   Instituto Leloir1, Buenos Aires, Argentina. We have conducted an unbiased genome-wide RNAi screen in Drosophila cells aimed to the identification of genes required for HIF dependent transcription. After 3 rounds of selection, 35 genes emerged as critical regulators of HIF activity in hypoxia, most of which had not been previously associated to HIF. We focused our analysis on components of chromatin remodeling complexes, including Reptin, Pontin Brahma and Moira, the transcription elongation factors Spt5 and Spt6 and the miRNA pathway component Argonaute 1. We have validated the requirement of these genes for the activation of a hypoxia inducible reporter in transgenic flies and by assessing the expression of 4 endogenous dHIF target genes in Drosophila embryos. Studies carried out both in cell culture and in vivo confirmed the physiological requirement of Argonaute 1 and the miRNA machinery for HIF dependent transcription.