The Hippo pathway promotes cell survival in response to chemical stress.

Cellular stress defense mechanisms have evolved to maintain homeostasis in response to a broad variety of environmental challenges. Stress signaling pathways activate multiple cellular programs that range from the activation of survival pathways to the initiation of cell death when cells are damaged beyond repair. To identify novel players acting in stress response pathways, we conducted a cell culture RNA interference (RNAi) screen using caffeine as a xenobiotic stress-inducing agent, as this compound is a well-established inducer of detoxification response pathways. Specifically, we examined how caffeine affects cell survival when Drosophila kinases and phosphatases were depleted via RNAi. Using this approach, we identified and validated 10 kinases and 4 phosphatases that are essential for cell survival under caffeine-induced stress both in cell culture and living flies. Remarkably, our screen yielded an enrichment of Hippo pathway components, indicating that this pathway regulates cellular stress responses. Indeed, we show that the Hippo pathway acts as a potent repressor of stress-induced cell death. Further, we demonstrate that Hippo activation is necessary to inhibit a pro-apoptotic program triggered by the interaction of the transcriptional co-activator Yki with the transcription factor p53 in response to a range of stress stimuli. Our in vitro and in vivo loss-of-function data therefore implicate Hippo signaling in the transduction of cellular survival signals in response to chemical stress.

Expressional and functional analysis of the male-specific cluster mst36F during Drosophila spermatogenesis.

mst36Fa and mst36Fb are two male-specific genes that are part of a novel gene family recently characterized in Drosophila melanogaster. The genes are strictly clustered and show an identical tissue and temporal expression pattern limited to the male germline. Here we demonstrate that the transcription of these two genes, which is triggered by different cis regulatory elements, responds to the same testisspecific factors encoded by the aly and can class meiotic arrest genes. RNA interference was used to decrease expression of these two genes. We obtained a reduction of fertility in the transgenic adult males compared to the wild type. These data suggest that the Mst36Fa and Mst36Fb proteins may have an important role in the production of functional sperm.

Structure and expression of a novel gene family showing male germline specific expression in Drosophila melanogaster.

We report the characterization of two novel genes of Drosophila melanogaster, named mst36Fa and mst36Fb. They define a novel gene family, showing identical time and tissue-specificity limited to male germ cells where their transcription starts during meiotic prophase. These two genes encode for two slightly basic proteins highly homologous to each other and fairly rich in leucine and glutamic acid. Although strictly clustered, these genes utilize different promoter regions as revealed by examination of transgenic flies bearing mst36F-promoter-lacZ reporter constructs and by reverse transcription-polymerase chain reaction assays. Our data suggest that at least one gene (mst36Fa) of the cluster is under translational repression until spermiogenesis suggesting a putative role in the spermatides differentiation. The present study is aimed at the structural analysis of these genes.