The ubiquitylome of developing cortical neurons

Menon, Shalini; Goldfarb, Dennis; Cousins, Emily; Major, Michael B.; Gupton, Stephanie L.

doi:10.1101/2020.10.13.337782

Cited by 3 publications

(4 citation statements)

References 13 publications

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“…It has been suggested that TRIM9 and TRIM67 can act in antagonistic ways, regulating VASP degradation and filopodia extensions (6). However, ubiquitylome analysis in mouse cortical neurons indicated overlapping and non-overlapping targets of TRIM9 and TRIM67, suggesting that both E3 ligases could have redundant and non-redundant functions (20). Our study shows an enrichment of TRIM9 protein levels, physical interaction and colocalization of TRIM9 with TRIM67.…”

Section: Discussionmentioning

confidence: 54%

“…The TRIM protein family also has E3 ubiquitin ligase activity, and direct ubiquitination targets of TRIM67 has so far been restricted to 80-K-H, such that TRIM67-induced degradation of 80-K-H leads to alteration of Ras signaling in neuroblastoma (10). A recent MS-based study detected potential TRIM67 and TRIM9 ubiquitination targets using mouse primary cortical neurons that need further functional characterization (20). In addition, our MS and co-IP-MS experiments using human glioma cell lines also provide datasets to further identify potential TRIM67 targets and interaction partners.…”

Section: Discussionmentioning

confidence: 99%

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TRIM67 drives tumorigenesis in oligodendrogliomas through Rho GTPase-dependent membrane blebbing

Demirdizen

Al‐Ali

Narayanan

et al. 2021

Preprint

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Oligodendrogliomas are a subtype of isocitrate dehydrogenase (IDH) mutant gliomas defined by the co-deletion of chromosome arms 1p and 19q. Although the somatic genomic alterations of oligodendrogliomas have been well described, transcriptional changes unique to these tumors are not well studied. Here, we identify Tripartite Motif Containing 67 (TRIM67), an E3 ubiquitin ligase with essential roles during neuronal development, as an oncogene distinctly upregulated in oligodendrogliomas. We characterize the function of TRIM67 using high throughput assays, including RNA sequencing, total lysate-mass spectrometry (MS) and co-immunoprecipitation (IP)-MS using human neural progenitor cells and patient-derived glioma tumorspheres constitutively overexpressing TRIM67. Our high throughput data suggest that TRIM67 overexpression alters the abundance of cytoskeletal proteins, which were validated by functional assays, including immunofluorescence (IF) staining, co-IP and western blotting (WB). Additionally, IF staining results indicate that TRIM67 ectopic expression induces formation of membrane blebs in glioma cells, which could be reverted with the nonmuscle class II myosin inhibitor blebbistatin and selective ROCK inhibitor fasudil. GTP pulldown and WB assays further indicate that Rho GTPase/ROCK2 signaling is altered upon TRIM67 ectopic expression. Phenotypically, TRIM67 expression resulted in higher cell motility in wound healing experiments, reduced cell adherence in adhesion assays, accelerated tumor growth and reduced survival in mouse orthotopic implantation models of an oligodendroglioma-derived patient tumorsphere line. Taken together, our results demonstrate that upregulated TRIM67 induces blebbing-based rounded cell morphology through Rho GTPase/ROCK-mediated signaling thereby contributing to glioma pathogenesis.SignificanceWe identify TRIM67 as a novel oncogene in oligodendroglioma that leads to increased cell motility, tumor growth, reduced adhesion, and survival in mice. Our results also show that constitutive TRIM67 expression transforms cell morphology from an adherent to a rounded appearance with membrane blebs. Mechanistic alteration of actin cytoskeleton and Rho GTPase signaling upon TRIM67 upregulation underlies the rounded cell structure and the membrane blebbing phenotype. TRIM67-induced blebbing is specifically regulated by RHOA-RAC1-ROCK2 signaling axis. TRIM67 overexpression also alters pathways associated with cell migration and wound healing in various glioma cell lines and human neural progenitor cells, suggesting a general oncogenic mechanism in gliomas. Overall, our study highlights TRIM67 as a novel player orchestrating cytoskeleton, Rho GTPase signaling and bleb-based cell movement, ultimately causing tumorigenic outcomes in oligodendrogliomas.

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Section: Discussionmentioning

confidence: 54%

Section: Discussionmentioning

confidence: 99%

TRIM67 drives tumorigenesis in oligodendrogliomas through Rho GTPase-dependent membrane blebbing

Demirdizen

Al‐Ali

Narayanan

et al. 2021

Preprint

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“…In particular, we have shown that TRIM9 directly interacts and ubiquitinates DCC (Plooster et al ., 2017), as well as the actin polymerase VASP (Menon et al ., 2015; Boyer et al ., 2020). Although our lab has worked to identify new substrates and interacting partners of TRIM9 during axon guidance (Menon et al ., 2020, 2021), there are still dozens of unvalidated candidates waiting to be explored.…”

Section: Discussionmentioning

confidence: 99%

The E3 ubiquitin ligase TRIM9 regulates synaptic function and actin dynamics

McCormick,

Evans,

Barker

et al. 2024

Preprint

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During neuronal development, dynamic filopodia emerge from dendrites and mature into functional dendritic spines during synaptogenesis. Dendritic filopodia and spines respond to extracellular cues, influencing dendritic spine shape and size as well as synaptic function. Previously, the E3 ubiquitin ligase TRIM9 was shown to regulate filopodia in early stages of neuronal development, including netrin-1 dependent axon guidance and branching. Here we demonstrate TRIM9 also localizes to dendritic filopodia and spines of murine cortical and hippocampal neurons during synaptogenesis and is required for synaptic responses to netrin-1. In particular, TRIM9 is enriched in the post-synaptic density (PSD) within dendritic spines and loss of Trim9 alters the PSD proteome, including the actin cytoskeleton landscape. While netrin-1 exposure induces accumulation of the Arp2/3 complex and filamentous actin in dendritic spine heads, this response is disrupted by genetic deletion ofTrim9. In addition, we document changes in synaptic receptors associated with loss ofTrim9. These defects converge on a loss of netrin-dependent increases in neuronal firing rates, indicating TRIM9 is required downstream of synaptic netrin-1 signaling. We propose TRIM9 regulates cytoskeletal dynamics in dendritic spines and is required for the proper response to synaptic stimuli.

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“…, 2020 ). Although our lab has worked to identify new substrates and interacting partners of TRIM9 during axon guidance ( Menon et al. , 2020 , 2021 ), there are still dozens of unvalidated candidates waiting to be explored.…”

Section: Discussionmentioning

confidence: 99%

The E3 ubiquitin ligase TRIM9 regulates synaptic function and actin dynamics in response to netrin-1

McCormick,

Evans,

Barker

et al. 2024

MBoC

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During neuronal development, dynamic filopodia emerge from dendrites and mature into functional dendritic spines during synaptogenesis. Dendritic filopodia and spines respond to extracellular cues, influencing dendritic spine shape and size as well as synaptic function. Previously, the E3 ubiquitin ligase TRIM9 was shown to regulate filopodia in early stages of neuronal development, including netrin-1 dependent axon guidance and branching. Here we demonstrate TRIM9 also localizes to dendritic filopodia and spines of murine cortical and hippocampal neurons during synaptogenesis and is required for synaptic responses to netrin. In particular, TRIM9 is enriched in the post-synaptic density (PSD) within dendritic spines and loss of Trim9 alters the PSD proteome, including the actin cytoskeleton landscape. While netrin exposure induces accumulation of the Arp2/3 complex and filamentous actin in dendritic spine heads, this response is disrupted by genetic deletion of Trim9. In addition, we document changes in the synaptic receptors associated with loss of Trim9. These defects converge on a loss of netrin-dependent increases in neuronal firing rates, indicating TRIM9 is required downstream of synaptic netrin-1 signaling. We propose TRIM9 regulates cytoskeletal dynamics in dendritic spines and is required for the proper response to synaptic stimuli.

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The ubiquitylome of developing cortical neurons

Cited by 3 publications

References 13 publications

TRIM67 drives tumorigenesis in oligodendrogliomas through Rho GTPase-dependent membrane blebbing

TRIM67 drives tumorigenesis in oligodendrogliomas through Rho GTPase-dependent membrane blebbing

The E3 ubiquitin ligase TRIM9 regulates synaptic function and actin dynamics

The E3 ubiquitin ligase TRIM9 regulates synaptic function and actin dynamics in response to netrin-1

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