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Bapat, J.

Publications and source records attributed to Bapat, J..

2 recordsLinked to original sources

Biomolecular condensates of Eya drive transcriptional co-activation associated with eye development

Eyes absent (Eya), a transcriptional coactivator essential for eye development in Drosophila, also plays an important role in organ development in mammals and is associated with several diseases. To better understand the mechanism of Eya-mediated transcriptional co-activation, we find that Eya forms biomolecular condensates with liquid-like properties. These condensates function as possible transcription hubs, through the compartmentalization of key eye development regulators, the transcription factors So, Dac, Optix, Ey, and the kinase Nemo, RNA Pol II machinery, P300/CBP, and show the presence of target RNAs. We map the PST-TPM-PST region of Eya, which is critical for transcriptional co-activation and eye development, to be crucial for condensate formation as well. We identify a human deafness-associated mutation mapping to a conserved Drosophila site in the PST-TPM-PST region, which impairs transcriptional co-activation and shifts the material property of the Eya condensates to a less dynamic state. Our findings thus provide evidence that the condensation of Eya and its dynamic state are integral to transcription regulation and its role in development and disease mechanisms.

cell biology↗

Neuronal Ndst1 depletion accelerates prion protein clearance and slows neurodegeneration in prion infection

Select prion diseases are characterized by widespread cerebral plaque-like deposits of amyloid fibrils enriched in heparan sulfate (HS), a major extracellular matrix component. HS facilitates fibril formation in vitro, yet how HS impacts fibrillar plaque growth within the brain is unclear. Here we found that prion-bound HS chains are highly sulfated, and that the sulfation is essential for HS accelerating prion conversion in vitro. Using conditional knockout mice to deplete the HS sulfation enzyme, Ndst1 (N-deacetylase, N-sulfotransferase), from neurons or astrocytes, we investigated how reducing HS sulfation impacts survival and prion aggregate distribution during a prion infection. Neuronal Ndst1-depleted mice survived longer and showed fewer and smaller parenchymal plaques, shorter fibrils, and increased vascular amyloid, consistent with enhanced aggregate transit toward perivascular drainage channels. The prolonged survival was strain-dependent, affecting mice infected with extracellular, plaque-forming, but not membrane bound, prion strains. Live PET imaging revealed rapid clearance of prion protein monomers into the CSF in mice expressing unsulfated HS, further suggesting that HS sulfate groups hinder transit of extracellular prion monomers. Our results directly show how a host cofactor slows the spread of prion protein through the extracellular space and identify an enzyme target to facilitate aggregate clearance. Author summaryPrions cause a rapidly progressive neurologic disease and death with no curative treatment available. Prion aggregates accumulate exponentially in the brain in affected individuals triggering neuronal loss and neuroinflammation. Yet the additional molecules that facilitate aggregation are largely unknown, and their identification may lead to new therapeutic targets. We have found that prions in the brain preferentially bind to a highly sulfated endogenous polysaccharide, known as heparan sulfate (HS). Here we use genetically modified mice that express poorly sulfated neuron-derived HS, and infect mice with different prions strains. We find that the mice infected with a plaque-forming prion strain show a prolonged survival and fewer plaques compared to the controls. We also found that the prion protein was efficiently transported in the interstitial fluid in mice having poorly sulfated HS, suggesting that the prion protein is more readily cleared from the brain. Our study provides insight into how HS retains prion aggregates in the brain to accelerate disease and indicates the specific HS biosynthetic enzymes to target for enhancing protein clearance.

pathology↗