Enhanced oligomerization of full-length RAGE by synergy of the interaction of its domains.

The pattern recognition receptor RAGE (receptor for advanced glycation end-products) transmits proinflammatory signals in several inflammation-related pathological states, including vascular diseases, cancer, neurodegeneration and diabetes. Its oligomerization is believed to be important in signal transduction, but RAGE oligomeric structures and stoichiometries remain unclear. Different oligomerization modes have been proposed in studies involving different truncated versions of the extracellular parts of RAGE. Here, we provide basic characterization of the oligomerization patterns of full-length RAGE (including the transmembrane (TM) and cytosolic regions) and compare the results with oligomerization modes of its four truncated fragments. For this purpose, we used native mass spectrometry, analytical ultracentrifugation, and size-exclusion chromatography coupled with multi-angle light scattering. Our results confirm known oligomerization tendencies of separate domains and highlight the enhanced oligomerization properties of full-length RAGE. Mutational analyses within the GxxxG motif of the TM region show sensitivity of oligomeric distributions to the TM sequence. Using hydrogen-deuterium exchange, we mapped regions involved in TM-dependent RAGE oligomerization. Our data provide experimental evidence for the major role of the C2 and TM domains in oligomerization, underscoring synergy among different oligomerization contact regions along the RAGE sequence. These results also explain the variability of obtained oligomerization modes in RAGE fragments

Valtion aluehallintovirastot ja niiden ylijohtajat: Pohjoiseurooppalainen analogia Ranskan prefeikteille

This chapter examines the closest Finnish analogy to the French function of the prefect. In Finland, since 2010, this function has been vested in the institution of the State Regional Administrative Agency (SRAA, aluehallintovirasto, ‘AVI’). There are six SRAAs, each headed by a Chief Director (ylijohtaja) nominated by the government. The study had four main findings. First, despite ambiguity in institutional terminology, classifications, boundaries and identities concerning the SRAA, one can discern few true functional or structural deficiencies. Second, the SRAA is a hybrid between an institution of its own and a territorial representative of either government ministries or government agencies, to which is related the fact that each SRAA has both responsibilities concerning its territory and nationwide responsibilities. Third, tensions between performance and institutional legitimation prevail in the institution of the SRAA, but again without serious deficiencies. Fourth, the 2010 substitution of the SRAA for the former Province comprised a radical institutional change. The 2015–2019 Finnish government intended to abolish the SRAAs, but the subsequent government abandoned that reform, and ultimately by mid-2020 it became clear that the institution of the SRAA was here to stay after all.Peer reviewe

New Public Management reforms in Europe and their effects: findings from a 20-country top executive survey

This article assesses the impact of New Public Management (NPM)-style reforms in European countries as perceived by top public sector officials. Using data from an executive survey conducted in 20 European countries, we look at the relationship between five key NPM reforms (downsizing, agencification, contracting out, customer orientation and flexible employment practices) and four dimensions of public sector performance: cost efficiency, service quality, policy coherence and coordination, and equal access to services. Structural equation modelling reveals that treating service users as customers and flexible employment are positively related to improvements on all four dimensions of performance. Contracting out and downsizing are both positively related to improved efficiency, but downsizing is also associated with worse service quality. The creation of autonomous agencies is unrelated to performance. This suggests that policy-makers seeking to modernize the public sector should prioritize managerial reforms within public organizations over structural transformations

Formation of Trans-Activation Competent HIV-1 Rev:RRE Complexes Requires the Recruitment of Multiple Protein Activation Domains

The HIV-1 Rev trans-activator is a nucleocytoplasmic shuttle protein that is essential for virus replication. Rev directly binds to unspliced and incompletely spliced viral RNA via the cis-acting Rev Response Element (RRE) sequence. Subsequently, Rev oligomerizes cooperatively and interacts with the cellular nuclear export receptor CRM1. In addition to mediating nuclear RNA export, Rev also affects the stability, translation and packaging of Rev-bound viral transcripts. Although it is established that Rev function requires the multimeric assembly of Rev molecules on the RRE, relatively little is known about how many Rev monomers are sufficient to form a trans-activation competent Rev:RRE complex, or which specific activity of Rev is affected by its oligomerization. We here analyzed by functional studies how homooligomer formation of Rev affects the trans-activation capacity of this essential HIV-1 regulatory protein. In a gain-of-function approach, we fused various heterologous dimerization domains to an otherwise oligomerization-defective Rev mutant and were able to demonstrate that oligomerization of Rev is not required per se for the nuclear export of this viral trans-activator. In contrast, however, the formation of Rev oligomers on the RRE is a precondition to trans-activation by directly affecting the nuclear export of Rev-regulated mRNA. Moreover, experimental evidence is provided showing that at least two protein activation domains are required for the formation of trans-activation competent Rev:RRE complexes. The presented data further refine the model of Rev trans-activation by directly demonstrating that Rev oligomerization on the RRE, thereby recruiting at least two protein activation domains, is required for nuclear export of unspliced and incompletely spliced viral RNA

Interrogating Membrane Protein Structure and Lipid Interactions by native mass spectrometry

Native mass spectrometry and native ion mobility mass spectrometry are now established techniques in structural biology, with recent work developing these methods for the study of integral membrane proteins reconstituted in both lipid bilayer and detergent environments. Here we show how native mass spectrometry can be used to interrogate integral membrane proteins, providing insights into conformation, oligomerization, subunit composition/stoichiometry, and interactions with detergents/lipids/drugs. Furthermore, we discuss the sample requirements and experimental considerations unique to integral membrane protein native mass spectrometry research

Coordinating for Cohesion in the Public Sector of the Future – final summary

status: publishe