@article {48069, title = {RNA-binding profiles of Drosophila CPEB proteins Orb and Orb2.}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, year = {2016}, month = {2016 Oct 24}, abstract = {

Localized protein translation is critical in many biological contexts, particularly in highly polarized cells, such as neurons, to regulate gene expression in a spatiotemporal manner. The cytoplasmic polyadenylation element-binding (CPEB) family of RNA-binding proteins has emerged as a key regulator of mRNA transport and local translation required for early embryonic development, synaptic plasticity, and long-term memory (LTM). Drosophila Orb and Orb2 are single members of the CPEB1 and CPEB2 subfamilies of the CPEB proteins, respectively. At present, the identity of the mRNA targets they regulate is not fully known, and the binding specificity of the CPEB2 subfamily is a matter of debate. Using transcriptome-wide UV cross-linking and immunoprecipitation, we define the mRNA-binding sites and targets of Drosophila CPEBs. Both Orb and Orb2 bind linear cytoplasmic polyadenylation element-like sequences in the 3{\textquoteright} UTRs of largely overlapping target mRNAs, with Orb2 potentially having a broader specificity. Both proteins use their RNA-recognition motifs but not the Zinc-finger region for RNA binding. A subset of Orb2 targets is translationally regulated in cultured S2 cells and fly head extracts. Moreover, pan-neuronal RNAi knockdown of these targets suggests that a number of these targets are involved in LTM. Our results provide a comprehensive list of mRNA targets of the two CPEB proteins in Drosophila, thus providing insights into local protein synthesis involved in various biological processes, including LTM.

}, issn = {1091-6490}, doi = {10.1073/pnas.1603715113}, author = {Stepien, Barbara Krystyna and Oppitz, Cornelia and Gerlach, Daniel and Dag, Ugur and Novatchkova, Maria and Kr{\"u}ttner, Sebastian and Stark, Alexander and Keleman, Krystyna} } @article {46887, title = {Synaptic Orb2A bridges memory acquisition and late memory consolidation in Drosophila.}, journal = {Cell Reports}, volume = {11}, year = {2015}, month = {2015 Jun 30}, pages = {1953-65}, abstract = {

To adapt to an ever-changing environment, animals consolidate some, but not all, learning experiences to long-term memory. In mammals, long-term memory consolidation often involves neural pathway reactivation hours after memory acquisition. It is not known whether this delayed-reactivation schema is\ common across the animal kingdom or how information is stored during the delay period. Here, we show\ that, during courtship suppression learning, Drosophila exhibits delayed long-term memory consolidation. We also show that the same class of dopaminergic neurons engaged earlier in memory acquisition is also both necessary and sufficient for delayed long-term memory consolidation. Furthermore, we present evidence that, during learning, the translational regulator Orb2A tags specific synapses of mushroom body neurons for later consolidation. Consolidation involves the subsequent recruitment of Orb2B and the activity-dependent synthesis of CaMKII. Thus, our results provide evidence for the role of a neuromodulated, synapse-restricted molecule bridging memory acquisition and long-term memory consolidation in a learning animal.

}, issn = {2211-1247}, doi = {10.1016/j.celrep.2015.05.037}, author = {Kr{\"u}ttner, Sebastian and Traunm{\"u}ller, Lisa and Dag, Ugur and Jandrasits, Katharina and Stepien, Barbara and Iyer, Nirmala and Fradkin, Lee G and Noordermeer, Jasprina N and Mensh, Brett D and Keleman, Krystyna} } @article {6854, title = {Dopamine neurons modulate pheromone responses in Drosophila courtship learning.}, journal = {Nature}, volume = {489}, year = {2012}, month = {2012 Sep 6}, pages = {145-9}, abstract = {

Learning through trial-and-error interactions allows animals to adapt innate behavioural {\textquoteleft}rules of thumb{\textquoteright} to the local environment, improving their prospects for survival and reproduction. Naive Drosophila melanogaster males, for example, court both virgin and mated females, but learn through experience to selectively suppress futile courtship towards females that have already mated. Here we show that courtship learning reflects an enhanced response to the male pheromone cis-vaccenyl acetate (cVA), which is deposited on females during mating and thus distinguishes mated females from virgins. Dissociation experiments suggest a simple learning rule in which unsuccessful courtship enhances sensitivity to cVA. The learning experience can be mimicked by artificial activation of dopaminergic neurons, and we identify a specific class of dopaminergic neuron that is critical for courtship learning. These neurons provide input to the mushroom body (MB) γ lobe, and the DopR1 dopamine receptor is required in MBγ neurons for both natural and artificial courtship learning. Our work thus reveals critical behavioural, cellular and molecular components of the learning rule by which Drosophila adjusts its innate mating strategy according to experience.

}, keywords = {Acetates, Animal, Animals, Brain, Courtship, Dopamine, Dopaminergic Neurons, Drosophila melanogaster, Female, Learning, Male, Mushroom Bodies, Oleic Acids, Pheromones, Presynaptic Terminals, Receptors, Sex Attractants, Sexual Behavior, Synaptic Transmission}, issn = {1476-4687}, doi = {10.1038/nature11345}, author = {Keleman, Krystyna and Vrontou, Eleftheria and Kr{\"u}ttner, Sebastian and Yu, Jai Y and Kurtovic-Kozaric, Amina and Dickson, Barry J} } @article {6856, title = {Drosophila CPEB Orb2A mediates memory independent of Its RNA-binding domain.}, journal = {Neuron}, volume = {76}, year = {2012}, month = {2012 Oct 18}, pages = {383-95}, abstract = {

Long-term memory and synaptic plasticity are thought to require the synthesis of new proteins at activated synapses. The CPEB family of RNA binding proteins, including Drosophila Orb2, has been implicated in this process. The precise mechanism by which these molecules regulate memory formation is however poorly understood. We used gene targeting and site-specific transgenesis to specifically modify the endogenous orb2 gene in order to investigate its role in long-term memory formation. We show that the Orb2A and Orb2B isoforms, while both essential, have distinct functions in memory formation. These two isoforms have common glutamine-rich and RNA-binding domains, yet Orb2A uniquely requires the former and Orb2B the latter. We further show that Orb2A induces Orb2 complexes in a manner dependent upon both its glutamine-rich region and neuronal activity. We propose that Orb2B acts as a conventional CPEB to regulate transport and/or translation of specific mRNAs, whereas Orb2A acts in an unconventional manner to form stable Orb2 complexes that are essential for memory to persist.

}, keywords = {Animals, Biogenic Amines, Brain, Cell Line, Chromatography, Courtship, Developmental, Drosophila, Drosophila Proteins, Embryo, Gene Expression Regulation, Genetically Modified, Genotype, Green Fluorescent Proteins, High Pressure Liquid, Immunoelectron, Immunoprecipitation, Larva, Learning, Male, Mass Spectrometry, Memory, Messenger, Microscopy, Mitogen-Activated Protein Kinases, mRNA Cleavage and Polyadenylation Factors, Mushroom Bodies, Mutation, Nonmammalian, Protein Isoforms, Protein Structure, RNA, RNA-Binding Proteins, Tertiary, Transcription Factors}, issn = {1097-4199}, doi = {10.1016/j.neuron.2012.08.028}, author = {Kr{\"u}ttner, Sebastian and Stepien, Barbara and Noordermeer, Jasprina N and Mommaas, Mieke A and Mechtler, Karl and Dickson, Barry J and Keleman, Krystyna} } @article {6858, title = {Function of the Drosophila CPEB protein Orb2 in long-term courtship memory.}, journal = {Nature Neuroscience}, volume = {10}, year = {2007}, month = {2007 Dec 28}, pages = {1587-93}, abstract = {

Both long-term behavioral memory and synaptic plasticity require protein synthesis, some of which may occur locally at specific synapses. Cytoplasmic polyadenylation element-binding (CPEB) proteins are thought to contribute to the local protein synthesis that underlies long-term changes in synaptic efficacy, but a role has not been established for them in the formation of long-term behavioral memory. We found that the Drosophila melanogaster CPEB protein Orb2 is acutely required for long-term conditioning of male courtship behavior. Deletion of the N-terminal glutamine-rich region of Orb2 resulted in flies that were impaired in their ability to form long-term, but not short-term, memory. Memory was restored by expressing Orb2 selectively in fruitless (fru)-positive gamma neurons of the mushroom bodies and by providing Orb2 function in mushroom bodies only during and shortly after training. Our data thus demonstrate that a CPEB protein is important in long-term memory and map the molecular, spatial and temporal requirements for its function in memory formation.

}, keywords = {Animal, Animals, Behavior, Computational Biology, Courtship, Drosophila, Drosophila Proteins, Female, Gene Expression, Genetically Modified, Green Fluorescent Proteins, Male, Memory, Memory Disorders, Mushroom Bodies, Neurons, Phylogeny, RNA-Binding Proteins, Sequence Deletion, Time Factors}, issn = {1097-6256}, doi = {10.1038/nn1996}, author = {Keleman, Krystyna and Kr{\"u}ttner, Sebastian and Alenius, Mattias and Dickson, Barry J} }