scholarly journals RBM45 associates with nuclear stress bodies and forms nuclear inclusions during chronic cellular stress and in neurodegenerative diseases

2019 ◽  
Author(s):  
Mahlon Collins ◽  
Yang Li ◽  
Robert Bowser
Keyword(s):  
Rna Binding ◽  
Cellular Stress ◽  
Heat Shock Factor 1 ◽  
Nuclear Inclusions ◽  
Nuclear Inclusion ◽  
Cell Type ◽  
Nuclear Speckles ◽  
Cytoplasmic Inclusions ◽  
Inclusion Formation ◽  
Self Association

AbstractRBM45 is a multifunctional RNA binding protein (RBP) found in cytoplasmic and nuclear inclusions in amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration (FTLD), and Alzheimer’s disease (AD). While cytoplasmic RBM45 inclusions contain other disease-associated proteins, nuclear RBM45 inclusions are morphologically and biochemically distinct from previously described nuclear inclusion pathology in these diseases. To better understand nuclear RBM45 aggregation and inclusion formation, we evaluated the association of RBM45 with a variety of membraneless nuclear organelles, including nuclear speckles, Cajal bodies, and nuclear gems. Under basal conditions, RBM45 is diffusely distributed throughout the nucleus and does not localize to a specific nuclear organelle. During cellular stress, however, the nuclear RBM45 distribution undergoes an RNA-binding dependent rearrangement wherein RBM45 coalesces into a small number of nuclear puncta. These puncta contain the nuclear stress body (NSB) markers heat shock factor 1 (HSF1) and scaffold attachment factor B (SAFB). During chronic stress, the persistent association of RBM45 with NSBs leads to the formation of large, insoluble nuclear RBM45 inclusions. RBM45 nuclear inclusions persist after stressor removal and NSB disassembly and the inclusions resemble the nuclear RBM45 pathology seen in ALS, FTLD, and AD. We also quantified the cell type- and disease-specific patterns of RBM45 pathology in ALS, FTLD, AD, and non-neurologic disease control subjects. RBM45 nuclear and cytoplasmic inclusions are found in neurons and glia in ALS, FTLD, and AD but not in controls. Across diseases, RBM45 nuclear inclusion pathology occurs more frequently than cytoplasmic RBM45 inclusion pathology and exhibits cell type-specific variation. Collectively, our results define new stress-associated functions of RBM45, a mechanism for its nuclear aggregation and inclusion formation, a role for NSBs in the pathogenesis of diseases such as ALS, FTLD, and AD, and further underscore the importance of self-association to both the normal and pathological functions of RBPs in these diseases.

scholarly journals Lysine acetylation regulates the RNA binding, subcellular localization and inclusion formation of FUS

2020 ◽  
Vol 29 (16) ◽  
pp. 2684-2697
Author(s):  
Alexandra Arenas ◽  
Jing Chen ◽  
Lisha Kuang ◽  
Kelly R Barnett ◽  
Edward J Kasarskis ◽  
...  
Keyword(s):  
Subcellular Localization ◽  
Histone Deacetylases ◽  
Rna Binding ◽  
Binding Protein ◽  
Nuclear Localization Sequence ◽  
Rna Recognition Motif ◽  
Creb Binding Protein ◽  
Cytoplasmic Inclusions ◽  
Inclusion Formation ◽  
Familial Als

Abstract Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the preferential death of motor neurons. Approximately 10% of ALS cases are familial and 90% are sporadic. Fused in sarcoma (FUS) is a ubiquitously expressed RNA-binding protein implicated in familial ALS and frontotemporal dementia (FTD). The physiological function and pathological mechanism of FUS are not well understood, particularly whether post-translational modifications play a role in regulating FUS function. In this study, we discovered that FUS was acetylated at lysine-315/316 (K315/K316) and lysine-510 (K510) residues in two distinct domains. Located in the nuclear localization sequence, K510 acetylation disrupted the interaction between FUS and Transportin-1, resulting in the mislocalization of FUS in the cytoplasm and formation of stress granule-like inclusions. Located in the RNA recognition motif, K315/K316 acetylation reduced RNA binding to FUS and decreased the formation of cytoplasmic inclusions. Treatment with deacetylase inhibitors also significantly reduced the inclusion formation in cells expressing ALS mutation P525L. More interestingly, familial ALS patient fibroblasts showed higher levels of FUS K510 acetylation as compared with healthy controls. Lastly, CREB-binding protein/p300 acetylated FUS, whereas both sirtuins and histone deacetylases families of lysine deacetylases contributed to FUS deacetylation. These findings demonstrate that FUS acetylation regulates the RNA binding, subcellular localization and inclusion formation of FUS, implicating a potential role of acetylation in the pathophysiological process leading to FUS-mediated ALS/FTD.

scholarly journals FUS inclusions disrupt RNA localization by sequestering kinesin-1 and inhibiting microtubule detyrosination

2017 ◽  
Vol 216 (4) ◽  
pp. 1015-1034 ◽  
Author(s):  
Kyota Yasuda ◽  
Sarah F. Clatterbuck-Soper ◽  
Meredith E. Jackrel ◽  
James Shorter ◽  
Stavroula Mili
Keyword(s):  
Rna Binding ◽  
Fibroblast Cell ◽  
Rna Localization ◽  
Cellular Functions ◽  
Cytoplasmic Inclusions ◽  
Inclusion Formation ◽  
Fused In Sarcoma ◽  
Lateral Sclerosis ◽  
Tubulin Carboxypeptidase ◽  
Dependent Mechanism

Cytoplasmic inclusions of the RNA-binding protein fused in sarcoma (FUS) represent one type of membraneless ribonucleoprotein compartment. Formation of FUS inclusions is promoted by amyotrophic lateral sclerosis (ALS)–linked mutations, but the cellular functions affected upon inclusion formation are poorly defined. In this study, we find that FUS inclusions lead to the mislocalization of specific RNAs from fibroblast cell protrusions and neuronal axons. This is mediated by recruitment of kinesin-1 mRNA and protein within FUS inclusions, leading to a loss of detyrosinated glutamate (Glu)–microtubules (MTs; Glu-MTs) and an inability to support the localization of RNAs at protrusions. Importantly, dissolution of FUS inclusions using engineered Hsp104 disaggregases, or overexpression of kinesin-1, reverses these effects. We further provide evidence that kinesin-1 affects MT detyrosination not through changes in MT stability, but rather through targeting the tubulin carboxypeptidase enzyme onto specific MTs. Interestingly, other pathological inclusions lead to similar outcomes, but through apparently distinct mechanisms. These results reveal a novel kinesin-dependent mechanism controlling the MT cytoskeleton and identify loss of Glu-MTs and RNA mislocalization as common outcomes of ALS pathogenic mutations.

scholarly journals An RNA-binding region regulates CTCF clustering and chromatin looping

2018 ◽  
Author(s):  
Anders S. Hansen ◽  
Tsung-Han S. Hsieh ◽  
Claudia Cattoglio ◽  
Iryna Pustova ◽  
Xavier Darzacq ◽  
...  
Keyword(s):  
Rna Binding ◽  
Embryonic Stem ◽  
Loop Formation ◽  
Cell Type ◽  
Binding Region ◽  
Chromatin Loops ◽  
Cell Type Specific ◽  
Self Association

Mammalian genomes are folded into Topologically Associating Domains (TADs), consisting of cell-type specific chromatin loops anchored by CTCF and cohesin. Since CTCF and cohesin are expressed ubiquitously, how cell-type specific CTCF-mediated loops are formed poses a paradox. Here we show RNase-sensitive CTCF self-association in vitro and that an RNA-binding region (RBR) mediates CTCF clustering in vivo. Intriguingly, deleting the RBR abolishes or impairs almost half of all chromatin loops in mouse embryonic stem cells. Disrupted loop formation correlates with abrogated clustering and diminished chromatin binding of the RBR mutant CTCF protein, which in turn results in a failure to halt cohesin-mediated extrusion. Thus, CTCF loops fall into at least 2 classes: RBR-independent and RBR-dependent loops. We suggest that evidence for distinct classes of RBR-dependent loops may provide a mechanism for establishing cell-specific CTCF loops regulated by RNAs and other RBR partner.

scholarly journals RBM45 Modulates the Antioxidant Response in Amyotrophic Lateral Sclerosis through Interactions with KEAP1

2015 ◽  
Vol 35 (14) ◽  
pp. 2385-2399 ◽  
Author(s):  
Nadine Bakkar ◽  
Arianna Kousari ◽  
Tina Kovalik ◽  
Yang Li ◽  
Robert Bowser
Keyword(s):  
Oxidative Stress ◽  
Spinal Cord ◽  
Amyotrophic Lateral Sclerosis ◽  
Motor Neurons ◽  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Antioxidant Response ◽  
Cytoplasmic Inclusions ◽  
Lateral Sclerosis

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the selective loss of motor neurons. Various factors contribute to the disease, including RNA binding protein dysregulation and oxidative stress, but their exact role in pathogenic mechanisms remains unclear. We have recently linked another RNA binding protein, RBM45, to ALS via increased levels of protein in the cerebrospinal fluid of ALS patients and its localization to cytoplasmic inclusions in ALS motor neurons. Here we show RBM45 nuclear exit in ALS spinal cord motor neurons compared to controls, a phenotype recapitulatedin vitroin motor neurons treated with oxidative stressors. We find that RBM45 binds and stabilizes KEAP1, the inhibitor of the antioxidant response transcription factor NRF2. ALS lumbar spinal cord lysates similarly show increased cytoplasmic binding of KEAP1 and RBM45. Binding of RBM45 to KEAP1 impedes the protective antioxidant response, thus contributing to oxidative stress-induced cellular toxicity. Our findings thus describe a novel link between a mislocalized RNA binding protein implicated in ALS (RBM45) and dysregulation of the neuroprotective antioxidant response seen in the disease.

scholarly journals Congregation of Orthopoxvirus Virions in Cytoplasmic A-Type Inclusions Is Mediated by Interactions of a Bridging Protein (A26p) with a Matrix Protein (ATIp) and a Virion Membrane-Associated Protein (A27p)

2010 ◽  
Vol 84 (15) ◽  
pp. 7592-7602 ◽  
Author(s):  
Amanda R. Howard ◽  
Andrea S. Weisberg ◽  
Bernard Moss
Keyword(s):  
Matrix Protein ◽  
Direct Interaction ◽  
Cowpox Virus ◽  
Cytoplasmic Inclusions ◽  
Virion Assembly ◽  
Inclusion Formation ◽  
Terminal Domain ◽  
Defective Mutant ◽  
Inclusion Protein ◽  
Type Inclusion

ABSTRACT Some orthopoxviruses, e.g., the cowpox, ectromelia, and raccoonpox viruses, form large, discrete cytoplasmic inclusions within which mature virions (MVs) are embedded by a process called occlusion. These inclusions, which may protect occluded MVs in the environment, are composed of aggregates of the A-type inclusion protein (ATIp), which is truncated in orthopoxviruses such as vaccinia virus (VACV) and variola virus that fail to form inclusions. In addition to an intact ATIp, occlusion requires the A26 protein (A26p). Although VACV contains a functional A26p, determined by complementation of a cowpox virus occlusion-defective mutant, its role in occlusion was unknown. We found that restoration of the full-length ATI gene was sufficient for VACV inclusion formation and the ensuing occlusion of MVs. A26p was present in inclusions even when virion assembly was inhibited, suggesting a direct interaction of A26p with ATIp. Analysis of a panel of ATIp mutants indicated that the C-terminal repeat region was required for inclusion formation and the N-terminal domain for interaction with A26p and occlusion. A26p is tethered to MVs via interaction with the A27 protein (A27p); A27p was not required for association of A26p with ATIp but was necessary for occlusion. In addition, the C-terminal domain of A26p, which mediates A26p-A27p interactions, was necessary but insufficient for occlusion. Taken together, the data suggest a model for occlusion in which A26p has a bridging role between ATIp and A27p, and A27p provides a link to the MV membrane.

scholarly journals RNA recognition and self-association of CPEB4 is mediated by its tandem RRM domains

2014 ◽  
Vol 42 (15) ◽  
pp. 10185-10195 ◽  
Author(s):  
Constanze Schelhorn ◽  
James M.B. Gordon ◽  
Lidia Ruiz ◽  
Javier Alguacil ◽  
Enrique Pedroso ◽  
...  
Keyword(s):  
Rna Binding ◽  
Titration Calorimetry ◽  
Rna Recognition Motif ◽  
Rna Recognition ◽  
Mobility Shift ◽  
Cytoplasmic Polyadenylation ◽  
C Terminus ◽  
Self Association ◽  
Mobility Shift Assay ◽  
A Minor

Abstract Cytoplasmic polyadenylation is regulated by the interaction of the cytoplasmic polyadenylation element binding proteins (CPEB) with cytoplasmic polyadenylation element (CPE) containing mRNAs. The CPEB family comprises four paralogs, CPEB1–4, each composed of a variable N-terminal region, two RNA recognition motif (RRM) and a C-terminal ZZ-domain. We have characterized the RRM domains of CPEB4 and their binding properties using a combination of biochemical, biophysical and NMR techniques. Isothermal titration calorimetry, NMR and electrophoretic mobility shift assay experiments demonstrate that both the RRM domains are required for an optimal CPE interaction and the presence of either one or two adenosines in the two most commonly used consensus CPE motifs has little effect on the affinity of the interaction. Both the single RRM1 and the tandem RRM1–RRM2 have the ability to dimerize, although representing a minor population. Self-association does not affect the proteins’ ability to interact with RNA as demonstrated by ion mobility–mass spectrometry. Chemical shift effects measured by NMR of the apo forms of the RRM1–RRM2 samples indicate that the two domains are orientated toward each other. NMR titration experiments show that residues on the β-sheet surface on RRM1 and at the C-terminus of RRM2 are affected upon RNA binding. We propose a model of the CPEB4 RRM1–RRM2–CPE complex that illustrates the experimental data.

scholarly journals Functional annotation of human long noncoding RNAs using chromatin conformation data

2021 ◽  
Author(s):  
Saumya Agrawal ◽  
Tanvir Alam ◽  
Masaru Koido ◽  
Ivan V. Kulakovskiy ◽  
Jessica Severin ◽  
...  
Keyword(s):  
Functional Annotation ◽  
Rna Binding ◽  
Functional Characterization ◽  
Cell Types ◽  
Chromatin Interaction ◽  
Spatial Proximity ◽  
Chromatin Conformation ◽  
Cell Type ◽  
Cell Type Specific ◽  
Rna Domains

AbstractTranscription of the human genome yields mostly long non-coding RNAs (lncRNAs). Systematic functional annotation of lncRNAs is challenging due to their low expression level, cell type-specific occurrence, poor sequence conservation between orthologs, and lack of information about RNA domains. Currently, 95% of human lncRNAs have no functional characterization. Using chromatin conformation and Cap Analysis of Gene Expression (CAGE) data in 18 human cell types, we systematically located genomic regions in spatial proximity to lncRNA genes and identified functional clusters of interacting protein-coding genes, lncRNAs and enhancers. Using these clusters we provide a cell type-specific functional annotation for 7,651 out of 14,198 (53.88%) lncRNAs. LncRNAs tend to have specialized roles in the cell type in which it is first expressed, and to incorporate more general functions as its expression is acquired by multiple cell types during evolution. By analyzing RNA-binding protein and RNA-chromatin interaction data in the context of the spatial genomic interaction map, we explored mechanisms by which these lncRNAs can act.

scholarly journals DIGESTIVE AND RESPIRATORY DISORDERS WITH INTRANUCLEAR INCLUSION BODIES IN TWO CALVES

1974 ◽  
Vol 25 (4) ◽  
pp. 5
Author(s):  
Χ. ΠΑΠΑΔΟΠΟΥΛΟΣ ◽  
Ε. ΣΙΜΟΣ
Keyword(s):  
Inclusion Bodies ◽  
The Other ◽  
Capillary Endothelium ◽  
Nuclear Inclusions ◽  
Nuclear Inclusion ◽  
Intranuclear Inclusion ◽  
Respiratory Disorders

The authors give a cytological description of liver and kidneys from two cal ves with digestive and respiratory troubles. Characteristic spherical nuclear inclusions were noticed. In one calf nuclear inclusion were noticed. In the other calf acidophilic nuclear inclusions were found in the kidneys •capillary endothelium especially in the cortex and partial vacuolated degeneration in the myocardial fibres.

scholarly journals Cellular stress induces cancer stem-like cells through expression of DNAJB8 by activation of heat shock factor 1

2018 ◽  
Vol 109 (3) ◽  
pp. 741-750 ◽  
Author(s):  
Hiroki Kusumoto ◽  
Yoshihiko Hirohashi ◽  
Satoshi Nishizawa ◽  
Masamichi Yamashita ◽  
Kazuyo Yasuda ◽  
...  
Keyword(s):  
Heat Shock ◽  
Cellular Stress ◽  
Heat Shock Factor ◽  
Heat Shock Factor 1

Cell Stress Signaling Cascades Regulating Cell Fate

2018 ◽  
Vol 24 (27) ◽  
pp. 3176-3183 ◽  
Author(s):  
Rohit Gundamaraju ◽  
Ravichandra Vemuri ◽  
Wai Chin Chong ◽  
Dominic P. Geraghty ◽  
Rajaraman Eri
Keyword(s):  
Cell Death ◽  
Cell Fate ◽  
Stress Responses ◽  
Cellular Stress ◽  
Cell Stress ◽  
Signaling Cascades ◽  
Stress Signaling ◽  
Cell Type ◽  
Induce Cell Death ◽  
Stressed Cell

Initiating anti-apoptotic signaling or triggering cell death depends to a great extent on the nature or source of cellular stress and cell type. Interplay between each stress response eventually determines the fate of stressed cell. Numerous factors induce cell death by a number of pathways including apoptosis, autophagy and necrosis. Not surprisingly, some of the pathways are interrelated to each other through a mediator that could articulate the entire mechanism. The present review attempts to consolidate all the pathways included in intrinsic cellular stress such as oxidative stress and autophagy, endoplasmic reticular stress (ERS) and mitophagy and apoptosis as fate in cell stress. These stress responses are a hallmark of numerous diseases including neurodegenerative diseases, diabetes and cancer. Understanding the cross-talk between different intrinsic cell stress responses will help to develop new therapeutic targets and hence lead to the development of new therapeutics.

Sign in / Sign up

Export Citation Format

Share Document