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Biology subjects

Mitchell, D. C.

Publications and source records attributed to Mitchell, D. C..

3 recordsLinked to original sources

Small molecule activation of the tumor suppressor kinase LKB1

The ability to identify and target oncogenic signals has transformed clinical oncology. Drug development for targeted therapies has historically focused on the inhibition of oncogenic kinases and GTPases. However, many cancer patients do not benefit from targeted approaches because their tumors lack targetable mutations. Therapeutic augmentation of tumor suppressive signaling could be a viable alternative but poses challenges. Specifically, designing compounds capable of stimulating kinase activity is more structurally challenging than inhibitor design, and most kinases lack targetable allosteric pockets. Inactivation of the liver kinase B1 (LKB1) tumor suppressor kinase is associated with poor prognosis and therapeutic resistance. Thus, augmented LKB1 function could be beneficial for cancer patients whose tumors retain intact copies of the gene. LKB1 signals as part of an obligate trimer including the scaffolding protein Mouse protein-25 (MO25) and the pseudokinase (PSK) STE20-related kinase adapter protein (STRAD). As STRAD binds but does not metabolize ATP, it provides the opportunity for a novel activation strategy. We have developed STRAD-binding compounds capable of activating LKB1 and demonstrate the therapeutic benefits of LKB1 activation in a target-dependent manner within cancer cell lines.

biochemistry↗

Highly specific intracellular ubiquitination of a small molecule

Ubiquitin is a small, highly conserved protein that acts as a posttranslational modification in eukaryotes. Ubiquitination of proteins frequently serves as a degradation signal, marking them for disposal by the proteasome. Here, we report a novel small molecule from a diversity-oriented synthesis library, BRD1732, that is directly ubiquitinated in cells, resulting in dramatic accumulation of inactive ubiquitin monomers and polyubiquitin chains causing broad inhibition of the ubiquitin-proteasome system. Ubiquitination of BRD1732 and its associated cytotoxicity are stereospecific and dependent upon two homologous E3 ubiquitin ligases, RNF19A and RNF19B. Our finding opens the possibility for indirect ubiquitination of a target through a ubiquitinated bifunctional small molecule, and more broadly raises the potential for posttranslational modification in trans.

biochemistry↗

The Hao-Fountain syndrome protein USP7 regulates neuronal connectivity in the brain via a novel p53-independent ubiquitin signaling pathway

Precise control of protein ubiquitination is essential for brain development, and hence, disruption of ubiquitin signaling networks can lead to neurological disorders. Mutations of the deubiquitinase USP7 cause the Hao-Fountain syndrome (HAFOUS), characterized by developmental delay, intellectual disability, autism, and aggressive behavior. Here, we report that conditional deletion of USP7 in excitatory neurons in the mouse forebrain triggers diverse phenotypes including sensorimotor deficits, learning and memory impairment, and aggressive behavior, resembling clinical features of HAFOUS. USP7 deletion induces neuronal apoptosis in a manner dependent of the tumor suppressor p53. However, most behavioral abnormalities in USP7 conditional mice persist despite p53 loss. Strikingly, USP7 deletion in the brain perturbs the synaptic proteome and dendritic spine morphogenesis independently of p53. Integrated proteomics analysis reveals that the neuronal USP7 interactome is enriched for proteins implicated in neurodevelopmental disorders and specifically identifies the RNA splicing factor Ppil4 as a novel neuronal substrate of USP7. Knockdown of Ppil4 in cortical neurons impairs dendritic spine morphogenesis, phenocopying the effect of USP7 loss on dendritic spines. These findings reveal a novel USP7-Ppil4 ubiquitin signaling link that regulates neuronal connectivity in the developing brain, with implications for our understanding of the pathogenesis of HAFOUS and other neurodevelopmental disorders.

neuroscience↗