Proteostasis Responses to Endogenous Alpha-Synuclein Aggregation in the Brain

α-Synuclein aggregation is implicated in several neurodegenerative diseases, including Parkinson’s disease (PD) and dementia with Lewy Bodies (DLB). Changes in cellular signaling pathways induced by this aggregation may contribute to cell death and disease pathogenesis. To investigate this, we used quantitative proteomics to measure the relative abundance changes of the proteome and phosphoproteome in response to aggregation of endogenous α-synuclein in the brain of a mouse model. Aggregation in this model is induced by the intrastriatal injection of α-synuclein pre-formed fibrils and recapitulates several cardinal features of human PD, including progressive aggregation concomitant with dopaminergic degeneration and motor symptoms. We quantified the relative abundance changes of 5,290 proteins and 2,763 phosphosites in wildtype mice and found significant changes in vesicle-mediated transport, RNA processing and the immune response. The immunoproteasome, an altered form of the constitutive proteasome that is induced in response to stress, was elevated in response to α-synuclein aggregation. Increased levels and activity of the immunoproteasome were found in human DLB compared with age-matched healthy controls. Additionally, the immunoproteasome degrades α-synuclein fibrils more efficiently than the constitutive proteasome. This is the first documented role of the immunoproteasome in synucleinopathies

Dynamic structural flexibility of α-synuclein

Abstractα-Synuclein is a conserved, abundantly expressed protein that is partially localized in pre-synaptic terminals in the central nervous system. The precise biological function(s) and structure of α-synuclein are under investigation. Recently, the native conformation and the presence of naturally occurring multimeric assemblies have come under debate. These are important deliberations because α-synuclein assembles into highly organized amyloid-like fibrils and non-amyloid amorphous aggregates that constitute the neuronal inclusions in Parkinson's disease and related disorders. Therefore understanding the nature of the native and pathological conformations is pivotal from the standpoint of therapeutic interventions that could maintain α-synuclein in its physiological state. In this review, we will discuss the existing evidence that define the physiological states of α-synuclein and highlight how the inherent structural flexibility of this protein may be important in health and disease

The Influence of Computational Mesh on the Prediction of Vortex Interactions about a Generic Missile Airframe

A research program has been underway for four years to study vortex interaction aerodynamics that are relevant to military air vehicle performance. The program has been conducted under the auspices of the NATO Science and Technology Organization (STO), Applied Vehicle Technology (AVT) panel by a Task Group with the identification of AVT-316. The Missile Facet of this group has concentrated their work on the vortical flow field around a generic missile airframe and its prediction via computational methods. This paper focuses on mesh-related effects and RANS simulations. Simulated vortex characteristics were found to depend strongly on the properties of the employed mesh, in terms of both resolution and topology. Predictions of missile aerodynamic coefficients show a great dependence on mesh properties as they are sensitive to computed vortex dynamics. Key suggestions about the desired mesh characteristics have been made. Based on these, a shared mesh was constructed to perform common analyses between the AVT-316 Missile Facet members. Mesh based uncertainties of the aerodynamic coefficient predictions were estimated via Richardson Extrapolation method

RNA-Binding Proteins in Amyotrophic Lateral Sclerosis and Neurodegeneration

Amyotrophic Lateral Sclerosis (ALS) is an adult onset neurodegenerative disease, which is universally fatal. While the causes of this devastating disease are poorly understood, recent advances have implicated RNA-binding proteins (RBPs) that contain predicted prion domains as a major culprit. Specifically, mutations in the RBPs TDP-43 and FUS can cause ALS. Cytoplasmic mislocalization and inclusion formation are common pathological features of TDP-43 and FUS proteinopathies. Though these RBPs share striking pathological and structural similarities, considerable evidence suggests that the ALS-linked mutations in TDP-43 and FUS can cause disease by disparate mechanisms. In a recent study, Couthouis et al. screened for protein candidates that were also involved in RNA processing, contained a predicted prion domain, shared other phenotypic similarities with TDP-43 and FUS, and identified TAF15 as a putative ALS gene. Subsequent sequencing of ALS patients successfully identified ALS-linked mutations in TAF15 that were largely absent in control populations. This study underscores the important role that perturbations in RNA metabolism might play in neurodegeneration, and it raises the possibility that future studies will identify other RBPs with critical roles in neurodegenerative disease

Induction of the Immunoproteasome Subunit Lmp7 Links Proteostasis and Immunity in α-Synuclein Aggregation Disorders

Accumulation of aggregated α-synuclein into Lewy bodies is thought to contribute to the onset and progression of dopaminergic neuron degeneration in Parkinson's disease (PD) and related disorders. Although protein aggregation is associated with perturbation of proteostasis, how α-synuclein aggregation affects the brain proteome and signaling remains uncertain. In a mouse model of α-synuclein aggregation, 6% of 6215 proteins and 1.6% of 8183 phosphopeptides changed in abundance, indicating conservation of proteostasis and phosphorylation signaling. The proteomic analysis confirmed changes in abundance of proteins that regulate dopamine synthesis and transport, synaptic activity and integrity, and unearthed changes in mRNA binding, processing and protein translation. Phosphorylation signaling changes centered on axonal and synaptic cytoskeletal organization and structural integrity. Proteostatic responses included a significant increase in the levels of Lmp7, a component of the immunoproteasome. Increased Lmp7 levels and activity were also quantified in postmortem human brains with PD and dementia with Lewy bodies. Functionally, the immunoproteasome degrades α-synuclein aggregates and generates potentially antigenic peptides. Expression and activity of the immunoproteasome may represent testable targets to induce adaptive responses that maintain proteome integrity and modulate immune responses in protein aggregation disorders. Keywords: Neurodegeneration, Parkinson's disease, Dopaminergic neurons, Immunoproteasome, Proteostasi