An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration

Flores, Brittany; Li, Xingli; Malik, Aatika; Martínez, José Luis Lázaro; Beg, Asim A.; Barmada, Sami J.

doi:10.1016/j.celrep.2019.03.093

Cited by 76 publications

(98 citation statements)

References 108 publications

(234 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Raw reads from murine frontal cortex and spinal cord (11)(12)(13), as well as human spinal cord, frontal cortex and cerebellum (11)(12)(13)(14)(15), were downloaded from Gene Expression Omnibus (GEO) with the SRA Toolkit v2.9.2. Reads were trimmed with TrimGalore v0.6.0 using automatic adapter detection and a minimum Phred score of 20.…”

Section: Rna Sequencingmentioning

confidence: 99%

Shortened TDP43 isoforms upregulated by neuronal hyperactivity drive TDP43 pathology in ALS

Weskamp¹,

Tank²,

Miguez³

et al. 2020

Journal of Clinical Investigation

Self Cite

100

101

View full text Add to dashboard Cite

Section: Rna Sequencingmentioning

confidence: 99%

Shortened TDP43 isoforms upregulated by neuronal hyperactivity drive TDP43 pathology in ALS

Weskamp¹,

Tank²,

Miguez³

et al. 2020

Journal of Clinical Investigation

Self Cite

100

101

View full text Add to dashboard Cite

“…We therefore tested this compound in neurons overexpressing TDP43, an RNA binding protein whose accumulation is integrally connected with both ALS and FTD 65, 66 . In prior studies, TDP43 overexpression reproduced characteristic features of disease, including the formation of ubiquitin- and TDP43-positive neuronal inclusions, cytoplasmic TDP43 mislocalization, and neurodegeneration 40, 67, 68 . As expected, TDP43 overexpression resulted in an increase in the risk of death in comparison to neurons transfected with EGFP, a control protein.…”

Section: Resultsmentioning

confidence: 81%

Uncovering active modulators of native macroautophagy through novel high-content screens

Safren

Tank

Santoro³

et al. 2019

Preprint

View full text Add to dashboard Cite

12Autophagy is an evolutionarily conserved pathway mediating the breakdown of cellular 13 proteins and organelles. Emphasizing its pivotal nature, autophagy dysfunction 14 contributes to many diseases; nevertheless, development of effective autophagy 15 modulating drugs is hampered by fundamental deficiencies in available methods for 16 measuring autophagic activity, or flux. To overcome these limitations, we introduced the 17 photoconvertible protein Dendra2 into the MAP1LC3B locus of human cells via 18 CRISPR/Cas9 genome editing, enabling accurate and sensitive assessments of 19 autophagy in living cells by optical pulse labeling. High-content screening of 1,500 tool 20 compounds provided construct validity for the assay and uncovered many new 21 autophagy modulators. In an expanded screen of 24,000 diverse compounds, we 22 identified additional hits with profound effects on autophagy. Further, the autophagy 23 activator NVP-BEZ235 exhibited significant neuroprotective properties in a 24 neurodegenerative disease model. These studies confirm the utility of the Dendra2-LC3 25 assay, while simultaneously highlighting new autophagy-modulating compounds that 26 display promising therapeutic effects. 27 28 Introduction 29 Macroautophagy (hereafter referred to as autophagy) is an essential pathway for 30 protein homeostasis whereby cytoplasmic proteins and organelles are delivered to 31 lysosomes for degradation 1 . Through the coordinated action of a series of autophagy-32 related (ATG) proteins and cargo receptors including p62/SQSTM1, NBR1, and 33 optinuerin 2 , substrates are sequestered within double membrane vesicles called 34 autophagosomes. Autophagosomes mature as they traffic along microtubules and 35 eventually fuse with lysosomes to form autolysosomes, wherein hydrolases degrade 36 autophagic cargo. The protein LC3 (ATG8) is an obligate component of autophagosome 37 membranes and is itself degraded within autolysosomes. For these reasons, it often 38 2 serves as both a marker of autophagosomes and as a representative autophagy 39 substrate 3 . 40Underscoring the critical requirement of autophagy in cellular homeostasis, 41 deletion of core autophagy genes in mice results in embryonic lethality 4,5,6 . Accordingly, 42 dysfunctional autophagy is linked to a wide spectrum of human disease including 43 neurodegeneration, cancer, metabolic disorders, infectious and cardiovascular 44 diseases 7 . Often these conditions involve deficiencies in one or more steps of 45 autophagy, resulting in impaired clearance of potentially toxic cellular components, 46 and/or a failure to replenish amino acids required for anabolic processes. In these 47 instances, enhancing the rate of autophagic cargo clearance, commonly referred to as 48 flux, would be beneficial. Conversely, autophagy can promote tumor progression and 49 resistance to chemotherapy for some cancers 8,9,10,11 . Here, autophagy inhibition may 50 represent a more apt therapeutic strategy 7 . 51Autophagy is of particular importance in the central nervous system...

show abstract

“…Given these data and the largely cytoplasmic localization of sTDP43, we surmised that sTDP43 accumulation would be toxic to mammalian neurons. We therefore utilized automated microscopy in conjunction with survival analysis to track individual neurons prospectively over time and determine their risk of death in an unbiased and highthroughput manner (14,15,59,69,70). Rodent primary mixed cortical neurons were transfected with mApple and EGFP-tagged TDP43 isoforms and imaged by fluorescence microscopy at 24h intervals for 10d (71).…”

Section: Stdp43 Overexpression Is Neurotoxicmentioning

confidence: 99%

“…Using available RNA-seq data obtained from human cell lines 59 , we identified two alternatively spliced TARDBP isoforms predicted to encode C-terminally truncated or shortened (s) TDP43 isoforms ( Figure 3A). Identical sTDP43 splice isoforms (TDP-S6 and TDP-S7) were detected in previous studies of TARDBP splicing 60,61 .…”

Section: Hyperexcitability Drives Tardbp Alternative Splicingmentioning

confidence: 99%

See 1 more Smart Citation

Shortened TDP43 isoforms upregulated by neuronal hyperactivity drive TDP43 pathology in ALS

Weskamp

Tank

Miguez

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

Cortical hyperexcitability and mislocalization of the RNA-binding protein TDP43 are highlyconserved features in amyotrophic lateral sclerosis (ALS). Nevertheless, the relationship between these phenomena remains poorly defined. Here, we showed that hyperexcitability recapitulates TDP43 pathology by upregulating shortened (s) TDP43 splice isoforms. These truncated isoforms accumulated in the cytoplasm and formed insoluble inclusions that sequestered full-length TDP43 via preserved N-terminal interactions. Consistent with these findings, sTDP43 overexpression was toxic to mammalian neurons, suggesting neurodegeneration arising from complementary gain-and loss-of-function mechanisms. In humans and mice, sTDP43 transcripts were enriched in vulnerable motor neurons, and we observed a striking accumulation of sTDP43 within neurons and glia of ALS patients. Collectively, these studies uncover a pathogenic role for alternative TDP43 isoforms in ALS, and implicate sTDP43 as a key contributor to the susceptibility of motor neurons in this disorder.

show abstract

An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration

Cited by 76 publications

References 108 publications

Shortened TDP43 isoforms upregulated by neuronal hyperactivity drive TDP43 pathology in ALS

Shortened TDP43 isoforms upregulated by neuronal hyperactivity drive TDP43 pathology in ALS

Uncovering active modulators of native macroautophagy through novel high-content screens

Shortened TDP43 isoforms upregulated by neuronal hyperactivity drive TDP43 pathology in ALS

Contact Info

Product

Resources

About