We propose to change the current approach to understanding the molecular basis of memory. Our approach challenges the current focus on quantitative identification of synaptic RNAs by developing new technologies to address protein synthesis-dependent synaptic plasticity: single- synapse-CLIP and synaptic translational profiling. These will allow us to redefine the problem from two new superimposed perspectives: the need to identify regulated RNA-protein complexes in specific synapses, and the need to define their role in translational regulation. Specific synapses will be studied: the apical dendrites of cerebellar Purkinje neurons (a site of motor learning), of CA1 pyramidal neurons in the stratum moleculare of the hippocampus (a site of associative memory), and of layer V pyramidal neurons of the visual cortex (a site of activity- dependent plasticity). Key RNA-protein complexes to be studied in these synapses will be Argonaute (Ago)-mRNA-miRNA ternary complexes, translationally regulated FMRP-mRNA complexes, and neuron-specific RNA regulatory protein-mRNA complexes known to be present in the dendrite and to bind 3'UTRs (nElavl (Hu proteins), Nova). These complexes will be compared with a delineation of all ribosome-mRNA synaptic complexes present in the same dendrites, allowing us to validate interactions by identifying translationally regulated synaptic mRNAs (synaptic translational profiling). Regulated dendritic RNAs will be further validated by assessing for their translational state in two well-studied paradigms of protein synthesis- dependent synaptic plasticity: that in the hippocampus, and in the visual cortex after dark rearng and subsequent light exposure. These studies will revolutionize our understanding of the nature and regulation of local synaptic mRNAs that underlie memory, setting the stage for new insight into neurologic diseases of memory such as Alzheimer's and other neurodegenerative diseases.
Dysregulation of RNA-the key molecule between genomic DNA and proteins-is increasingly recognized to lie at the root of human neurologic disease. More precisely, memory, and complex cognitive function in general, is thought to depend on the regulation of protein synthesis at the sites of neuronal connections. Understanding the mechanisms governing this regulation will be studied here in an entirely innovative set of experiments, and they hold the key to understanding disorders of memory and cognition.
|Ke, Shengdong; Pandya-Jones, Amy; Saito, Yuhki et al. (2017) m6A mRNA modifications are deposited in nascent pre-mRNA and are not required for splicing but do specify cytoplasmic turnover. Genes Dev 31:990-1006|
|Hwang, Hun-Way; Saito, Yuhki; Park, Christopher Y et al. (2017) cTag-PAPERCLIP Reveals Alternative Polyadenylation Promotes Cell-Type Specific Protein Diversity and Shifts Araf Isoforms with Microglia Activation. Neuron 95:1334-1349.e5|
|Korb, Erica; Herre, Margaret; Zucker-Scharff, Ilana et al. (2017) Excess Translation of Epigenetic Regulators Contributes to Fragile X Syndrome and Is Alleviated by Brd4 Inhibition. Cell 170:1209-1223.e20|
|Linderman, Jessica A; Kobayashi, Mariko; Rayannavar, Vinayak et al. (2017) Immune Escape via a Transient Gene Expression Program Enables Productive Replication of a Latent Pathogen. Cell Rep 18:1312-1323|
|Luna, Joseph M; Barajas, Juan M; Teng, Kun-Yu et al. (2017) Argonaute CLIP Defines a Deregulated miR-122-Bound Transcriptome that Correlates with Patient Survival in Human Liver Cancer. Mol Cell 67:400-410.e7|
|Scheckel, Claudia; Drapeau, Elodie; Frias, Maria A et al. (2016) Regulatory consequences of neuronal ELAV-like protein binding to coding and non-coding RNAs in human brain. Elife 5:|
|Leggere, Janelle C; Saito, Yuhki; Darnell, Robert B et al. (2016) NOVA regulates Dcc alternative splicing during neuronal migration and axon guidance in the spinal cord. Elife 5:|
|Saito, Yuhki; Miranda-Rottmann, Soledad; Ruggiu, Matteo et al. (2016) NOVA2-mediated RNA regulation is required for axonal pathfinding during development. Elife 5:|
|Scheel, Troels K H; Luna, Joseph M; Liniger, Matthias et al. (2016) A Broad RNA Virus Survey Reveals Both miRNA Dependence and Functional Sequestration. Cell Host Microbe 19:409-23|
|Hwang, Hun-Way; Park, Christopher Y; Goodarzi, Hani et al. (2016) PAPERCLIP Identifies MicroRNA Targets and a Role of CstF64/64tau in Promoting Non-canonical poly(A) Site Usage. Cell Rep 15:423-35|
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