bioRxiv Science⌕ Search

Biology subjects

Ramachandran, K. V.

Publications and source records attributed to Ramachandran, K. V..

2 recordsLinked to original sources

Spatial coupling of endogenous Tau translation and degradation by neuroproteasomes in dendrites revealed by STARFISH

Cells regulate protein synthesis, folding, and degradation to maintain proteostasis, and disruptions in these processes have been linked to neurodegenerative diseases. In Alzheimers disease (AD), the protein Tau mislocalizes from axons to the somatodendritic compartment and aggregates into pathological filaments. Although Tau aggregation is a hallmark of AD, the subcellular dynamics of its synthesis and degradation are not well characterized. Because nascent polypeptides are particularly susceptible to misfolding, local control of Tau synthesis and degradation may be essential to prevent aggregation. Here, we develop STARFISH, a method for visualizing the subcellular site of endogenous mRNA translation in primary neurons and in vivo with single-molecule sensitivity and near-codon resolution, without modifying the nascent polypeptide. Using STARFISH, we show that despite the broad distribution of Mapt mRNA, Tau is translated almost exclusively in neuronal dendrites, revealing an unexpected level of spatial regulation. We further identify that one-third of newly synthesized Tau is co- or peri-translationally degraded in dendrites by a neuronal-specific plasma membrane-associated proteasome, the neuroproteasome. Failure of neuroproteasome-mediated degradation leads to the protein synthesis-dependent accumulation of somatodendritically mislocalized Tau aggregates. These findings define a previously unrecognized proteostasis mechanism that counterbalances the constitutive physiological overproduction of Tau. We speculate that failure of this proteostasis system contributes directly to Tau aggregation in dendrites, defining a new pathomechanism in Alzheimers disease.

neuroscience↗

ApoE isoforms differentially regulate neuronal membrane proteasomes to shift the threshold for pathological aggregation of endogenous Tau

Neuroproteasomes are a subset of 20S proteasomes that are localized to the neuronal plasma membrane and degrade newly synthesized proteins. To date, the molecular composition of neuroproteasomes is undefined, and moreover, whether neuroproteasomes can influence protein aggregation with relevance to neurodegenerative disorders remains unexplored. Using a Cre-dependent conditional knock-in mouse line to endogenously tag the proteasome, we find that neuroproteasomes co-purify with ApoE, the most significant risk factor for late-onset Alzheimers Disease (AD). We discover that neuroproteasome membrane localization is differentially modulated by ApoE isoforms (E4<E3<E2) in vitro, in vivo, and in human postmortem samples. We synthesized selective, neuroproteasome-specific inhibitors and discovered that neuroproteasome inhibition induces aggregation of endogenous mouse and human Tau, without the need for seeding or pathogenic mutations. Using hApoE-KI/hTau-KI crosses, we find that ApoE isoforms differentially shift the aggregation threshold for Tau. Neuroproteasome inhibition in vivo is sufficient to induce sarkosyl-insoluble and Thioflavin-S positive endogenous Tau aggregates in only three days, which are completely abrogated by co-application of cycloheximide. Newly synthesized Tau levels increase threefold after neuroproteasome inhibition, leading us to posit that newly synthesized Tau is uniquely susceptible to aggregation due to neuroproteasome dysfunction. Overall, our data define neuroproteasomes as a pivotal proteostasis mechanism underlying the formation of endogenous Tau aggregates, which is directly regulated by the largest genetic risk factor for late-onset Alzheimers Disease.

neuroscience↗