Nucleocapsid protein of SARS-CoV-2 phase separates into RNA-rich polymerase-containing condensates

The etiologic agent of the Covid-19 pandemic is the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The viral membrane of SARS-CoV-2 surrounds a helical nucleocapsid in which the viral genome is encapsulated by the nucleocapsid protein. The nucleocapsid protein of SARS-CoV-2 is produced at high levels within infected cells, enhances the efficiency of viral RNA transcription, and is essential for viral replication. Here, we show that RNA induces cooperative liquid–liquid phase separation of the SARS-CoV-2 nucleocapsid protein. In agreement with its ability to phase separate in vitro, we show that the protein associates in cells with stress granules, cytoplasmic RNA/protein granules that form through liquid-liquid phase separation and are modulated by viruses to maximize replication efficiency. Liquid–liquid phase separation generates high-density protein/RNA condensates that recruit the RNA-dependent RNA polymerase complex of SARS-CoV-2 providing a mechanism for efficient transcription of viral RNA. Inhibition of RNA-induced phase separation of the nucleocapsid protein by small molecules or biologics thus can interfere with a key step in the SARS-CoV-2 replication cycle

VARIATIONS OF CLIMATE PARAMETERS AND THEIR IMPACT ON CABERNET SAUVIGNON AND SAUVIGNON BLANC PHENOLOGY IN CONDITIONS OF CENTRAL SERBIA

Research carried out in vineyard of King Peter I Karadjordjevic-Royal Winery at Oplenac-Topola municipality, on Sauvignon blanc and Cabernet sauvignon. Vineyard planted with Sauvignon blanc is geographically positioned at GPS coordinates N 44° 14' 4" and E 20° 41' 15" and Cabernet sauvignon is geographically positioned at GPS coordinates N 44° 14' 35" and E 20° 41' 22". Climat parameters (series from 1982-2011 year) included following parameters: mean monthly, vegetation (april-october) and annual temperature, active and effective temperature, precipitation distribution (annual and vegetation) and wind direction. Phenological observation included beginning and end of following phenophases: bleeding, budbreakt, shoot growing, flowering, berry development and ripening. The greatest variation Sauvignon blanc manifested in duration of grape ripening which is in 2010. lasted 46 days while in 2011. lasted 34 days. Cabernet sauvignon in 2011. had a lower number of days that have passed from bleeding to full maturity (209 days) compared to 2010., when it passed 217 days

Molecular interactions of FG nucleoporin repeats at high resolution

Proteins that contain repeat phenylalanine-glycine (FG) residues phase separate into oncogenic transcription factor condensates in malignant leukaemias, form the permeability barrier of the nuclear pore complex and mislocalize in neurodegenerative diseases. Insights into the molecular interactions of FG-repeat nucleoporins have, however, remained largely elusive. Using a combination of NMR spectroscopy and cryoelectron microscopy, we have identified uniformly spaced segments of transient β-structure and a stable preformed α-helix recognized by messenger RNA export factors in the FG-repeat domain of human nucleoporin 98 (Nup98). In addition, we have determined at high resolution the molecular organization of reversible FG–FG interactions in amyloid fibrils formed by a highly aggregation-prone segment in Nup98. We have further demonstrated that amyloid-like aggregates of the FG-repeat domain of Nup98 have low stability and are reversible. Our results provide critical insights into the molecular interactions underlying the self-association and phase separation of FG-repeat nucleoporins in physiological and pathological cell activities

Phosphorylation but Not Oligomerization Drives the Accumulation of Tau with Nucleoporin Nup98

Tau is a neuronal protein that stabilizes axonal microtubules (MTs) in the central nervous system. In Alzheimer’s disease (AD) and other tauopathies, phosphorylated Tau accumulates in intracellular aggregates, a pathological hallmark of these diseases. However, the chronological order of pathological changes in Tau prior to its cytosolic aggregation remains unresolved. These include its phosphorylation and detachment from MTs, mislocalization into the somatodendritic compartment, and oligomerization in the cytosol. Recently, we showed that Tau can interact with phenylalanine-glycine (FG)-rich nucleoporins (Nups), including Nup98, that form a diffusion barrier inside nuclear pore complexes (NPCs), leading to defects in nucleocytoplasmic transport. Here, we used surface plasmon resonance (SPR) and bio-layer interferometry (BLI) to investigate the molecular details of Tau:Nup98 interactions and determined how Tau phosphorylation and oligomerization impact the interactions. Importantly, phosphorylation, but not acetylation, strongly facilitates the accumulation of Tau with Nup98. Oligomerization, however, seems to inhibit Tau:Nup98 interactions, suggesting that Tau-FG Nup interactions occur prior to oligomerization. Overall, these results provide fundamental insights into the molecular mechanisms of Tau-FG Nup interactions within NPCs, which might explain how stress-and disease-associated posttranslational modifications (PTMs) may lead to Tau-induced nucleocytoplasmic transport (NCT) failure. Intervention strategies that could rescue Tau-induced NCT failure in AD and tauopathies will be further discussed

The TEEBAgriFood theory of change: from information to action

KEY MESSAGES • Information alone often fails to motivate change. Manipulation of data has led consumers to doubt scientific results, serving special interests at the expense of public benefit. Information overload implies the need for synthesis to enable better access and impact. • Rationalizations against the need for change include: fatalism, arguing that business is already changing of its own accord, that cheap food is more important than good food, and that the marketplace will adjust for externalities. • These views do not address the long-term systemic consequences of the global corporate model of food systems in a society that derives calories from corn syrup and protein from hamburger resulting in obesity and disease. • Free market, neoliberal policies are incapable of resolving externalities that affect public goods such as ecosystem services. Faith in the infallibility of the market is a shortcoming of mainstream economics. • Path dependency is a key barrier to change in food systems, causing inertia, but may also lock-in positive systemic change. A science of intentional systemic change is arising, grounded in better understanding of human economic behavior as the basis for collective action. • We espouse not one theory but rather a range of actor-relevant theories of change. • Consumer advocacy can bring businesses to assume greater responsibility for the effects of their actions. This theory of change has found expression in the threat of boycotts and reputational risk. • Certification has led to improvement in production practice within market niches but its true success begins when it pressures change in policy and practice throughout supply chains. • Governance of intentional transformation in food systems requires knowledge of political pressure points, and systematic efforts to shape narratives of principal actors, to redirect financial resources and to promote institutional and societal learning and adaptation. • We address the potential of multilateral organizations and agreements, national governments, the financial industry, agribusiness, producers and consumer groups to respond to the need for change. The roles of different actors are interlocking: there is no single point of entry for a theory of change. • The roles of principal actors are drawn along a continuum of change, suggesting specific roles and types of actions to be addressed in evaluation and intervention. Given societal concern, agents for change may persevere within government, agribusiness or civil society organizations; their ability to bring change is dynamic and opportunistic, and driven by strategic alliances. As levers of agrifood system transformation, it is crucial to engage influential governmental actors as change agents. • Actors’ respective ability to adopt the results of TEEBAgriFood studies as a tool to direct change will depend on how well those results are communicated and adopted as narratives by influential actors and as entry points for education and consumer consciousness

Sensory Communication

Contains table of contents for Section 2, an introduction, reports on nine research projects and a list of publications.National Institutes of Health Grant 5 R01 DC00117National Institutes of Health Grant 2 R01 DC00270National Institutes of Health Grant 1 P01 DC00361National Institutes of Health Grant 2 R01 DC00100National Institutes of Health Grant FV00428National Institutes of Health Grant 5 R01 DC00126U.S. Air Force - Office of Scientific Research Grant AFOSR 90-200U.S. Navy - Office of Naval Research Grant N00014-90-J-1935National Institutes of Health Grant 5 R29 DC0062