Filament assembly of the C. elegans lamin in the absence of helix 1A

Lamins are the major constituent of the nuclear lamina, a protein meshwork underlying the inner nuclear membrane. Nuclear lamins are type V intermediate filaments that assemble into ~3.5 nm thick filaments. To date, only the conditions for the in vitro assembly of Caenorhabditis elegans lamin (Ce-lamin) are known. Here, we investigated the assembly of Ce-lamin filaments by cryo-electron microscopy and tomography. We show that Ce-lamin is composed of ~3.5 nm protofilaments that further interact in vitro and are often seen as 6-8 nm thick filaments. We show that the assembly of lamin filaments is undisturbed by the removal of flexible domains, that is, the intrinsically unstructured head and tail domains. In contrast, much of the coiled-coil domains are scaffold elements that are essential for filament assembly. Moreover, our results suggest that Ce-lamin helix 1A has a minor scaffolding role but is important to the lateral assembly regulation of lamin protofilaments. Keywords: C. elegans; cryo-electron tomography; intermediate filaments; lamin

Güterabwägung in Zeiten der Covid-19-Pandemie

Zusammenfassung: Die Covid-19-Pandemie und der politische Umgang mit ihr beeinflussen sämtliche Bereiche des gesellschaftlichen Lebens auf vielfältige Weise. Ziel dieses Artikels ist es, eine Güterabwägung am Beispiel schwerkranker Kinder und ihrer Familien als vulnerabler Gruppe durchzuführen, um politische Entscheidungen ethisch zu reflektieren. Dafür wird herausgearbeitet, was die Kinderpalliativmedizin im Vergleich zu anderen medizinischen Fachbereichen auszeichnet, und dargestellt, dass ihr Fokus auf der Linderung des „Total Pain“ nach Cicely Saunders liegt. Anhand einer Kasuistik wird ersichtlich, dass die Kinderpalliativmedizin aus diesem Grund durch die vielfältigen Einflüsse der Pandemie auf die physische, psychische, soziale und spirituelle Dimension betreuter Familien vor besondere Herausforderungen gestellt wird. In diesem Zuge wird ebenfalls erläutert, dass Familien mit kinderpalliativmedizinischem Bedarf im Kontext der Covid-19-Pandemie als vulnerable Gruppe angesehen werden können und weshalb die Notwendigkeit einer ausführlichen Güterabwägung besteht. Im Rahmen einer anschließenden ethischen Diskussion wird eine derartige Güterabwägung unter der Einschränkung von Unsicherheit durchgeführt und auf die Problematik der politischen Situation als eines „tragischen Dilemmas“ eingegangen

Structure of hibernating ribosomes studied by cryoelectron tomography in vitro and in situ

CryoET shows the configuration of the ephemeral translationally inactive 100S ribosomal dimer

Karyopherins regulate nuclear pore complex barrier and transport function

Nucleocytoplasmic transport is sustained by karyopherins (Kaps) and a Ran guanosine triphosphate (RanGTP) gradient that imports nuclear localization signal (NLS)–specific cargoes (NLS-cargoes) into the nucleus. However, how nuclear pore complex (NPC) barrier selectivity, Kap traffic, and NLS-cargo release are systematically linked and simultaneously regulated remains incoherent. In this study, we show that Kap α facilitates Kap β 1 turnover and occupancy at the NPC in a RanGTP-dependent manner that is directly coupled to NLS-cargo release and NPC barrier function. This is underpinned by the binding affinity of Kap β 1 to phenylalanine–glycine nucleoporins (FG Nups), which is comparable with RanGTP·Kap β 1, but stronger for Kap α ·Kap β 1. On this basis, RanGTP is ineffective at releasing standalone Kap β 1 from NPCs. Depleting Kap α ·Kap β 1 by RanGTP further abrogates NPC barrier function, whereas adding back Kap β 1 rescues it while Kap β 1 turnover softens it. Therefore, the FG Nups are necessary but insufficient for NPC barrier function. We conclude that Kaps constitute integral constituents of the NPC whose barrier, transport, and cargo release functionalities establish a continuum under a mechanism of Kap-centric control

Marker-free image registration of electron tomography tilt-series

<p>Abstract</p> <p>Background</p> <p>Tilt series are commonly used in electron tomography as a means of collecting three-dimensional information from two-dimensional projections. A common problem encountered is the projection alignment prior to 3D reconstruction. Current alignment techniques usually employ gold particles or image derived markers to correctly align the images. When these markers are not present, correlation between adjacent views is used to align them. However, sequential pairwise correlation is prone to bias and the resulting alignment is not always optimal.</p> <p>Results</p> <p>In this paper we introduce an algorithm to find regions of the tilt series which can be tracked within a subseries of the tilt series. These regions act as landmarks allowing the determination of the alignment parameters. We show our results with synthetic data as well as experimental cryo electron tomography.</p> <p>Conclusion</p> <p>Our algorithm is able to correctly align a single-tilt tomographic series without the help of fiducial markers thanks to the detection of thousands of small image patches that can be tracked over a short number of images in the series.</p

Pore timing:the evolutionary origins of the nucleus and nuclear pore complex

The name “eukaryote” is derived from Greek, meaning “true kernel”, and describes the domain of organisms whose cells have a nucleus. The nucleus is thus the defining feature of eukaryotes and distinguishes them from prokaryotes (Archaea and Bacteria), whose cells lack nuclei. Despite this, we discuss the intriguing possibility that organisms on the path from the first eukaryotic common ancestor to the last common ancestor of all eukaryotes did not possess a nucleus at all—at least not in a form we would recognize today—and that the nucleus in fact arrived relatively late in the evolution of eukaryotes. The clues to this alternative evolutionary path lie, most of all, in recent discoveries concerning the structure of the nuclear pore complex. We discuss the evidence for such a possibility and how this impacts our views of eukaryote origins and how eukaryotes have diversified subsequent to their last common ancestor