LIVING FOSSIL RECORD OF THE PRIMORDIAL ACTIN CYTOSKELETON IN PRESENT-DAY ASGARD ARCHAEA

Ph.DDOCTOR OF PHILOSOPHY (SOM

The eukaryotic-like characteristics of small GTPase, roadblock and TRAPPC3 proteins from Asgard archaea

Membrane-enclosed organelles are defining features of eukaryotes in distinguishing these organisms from prokaryotes. Specification of distinct membranes is critical to assemble and maintain discrete compartments. Small GTPases and their regulators are the signaling molecules that drive membrane-modifying machineries to the desired location. These signaling molecules include Rab and Rag GTPases, roadblock and longin domain proteins, and TRAPPC3-like proteins. Here, we take a structural approach to assess the relatedness of these eukaryotic-like proteins in Asgard archaea, the closest known prokaryotic relatives to eukaryotes. We find that the Asgard archaea GTPase core domains closely resemble eukaryotic Rabs and Rags. Asgard archaea roadblock, longin and TRAPPC3 domain-containing proteins form dimers similar to those found in the eukaryotic TRAPP and Ragulator complexes. We conclude that the emergence of these protein architectures predated eukaryogenesis, however further adaptations occurred in proto-eukaryotes to allow these proteins to regulate distinct internal membranes

The effects of needle deformation during lumbar puncture

Objectives: The aim of this study is to assess deformation of the tip and deflection from the axis of 22-gauge Quincke needles when they are used for diagnostic lumbar puncture (LP). Thus, it can be determined whether constructional alterations of needles are important for predicting clinical problems after diagnostic LP. Materials and Methods: The 22-gauge Quincke needles used for diagnostic LP were evaluated. A specially designed protractor was used for measurement and evaluation. Waist circumference was measured in each patient. Patients were questioned about headaches occurring after LP. Results: A total of 115 Quincke-type spinal needles used in 113 patients were evaluated. No deflection was detected in 38 (33.1%) of the needles. Deflection between 0.1° and 5° occurred in 43 (37.3%) of the needles and deflection ≥ 5.1° occurred in 34 patients (29.6%). Forty-seven (41.5%) patients experienced post lumbar puncture headache (PLPH) and 13 (11.5%) patients experienced intracranial hypotension (IH). No statistically significant correlation between the degree of deflection and headache was found (P > 0.05). Epidural blood patch was performed for three patients. Deformity in the form of bending like a hook occurred in seven needles and IH occurred in six patients using these needles. Two of the needles used in three patients requiring blood patch were found to be bent. Conclusion: Deformation of needles may increase complications after LP. Needle deformation may lead to IH. In case of deterioration in the structure of the needle, termination of the puncture procedure and the use of a new needle could reduce undesirable clinical consequences, especially IH

Insights into the evolution of regulated actin dynamics via characterization of primitive gelsolin/cofilin proteins from Asgard archaea

International audienceAsgard archaea genomes contain potential eukaryotic-like genes that provide intriguing insight for the evolution of eukaryotes. The eukaryotic actin polymerization/depolymerization cycle is critical for providing force and structure in many processes, including membrane remodeling. In general, Asgard genomes encode two classes of actin-regulating proteins from sequence analysis, profilins and gelsolins. Asgard profilins were demonstrated to regulate actin filament nucleation. Here, we identify actin filament severing, capping, annealing and bundling, and monomer sequestration activities by gelsolin proteins from Thorarchaeota (Thor), which complete a eukaryotic-like actin depolymerization cycle, and indicate complex actin cytoskeleton regulation in Asgard organisms. Thor gelsolins have homologs in other Asgard archaea and comprise one or two copies of the prototypical gelsolin domain. This appears to be a record of an initial preeukaryotic gene duplication event, since eukaryotic gelsolins are generally comprise three to six domains. X-ray structures of these proteins in complex with mammalian actin revealed similar interactions to the first domain of human gelsolin or cofilin with actin. Asgard two-domain, but not one-domain, gelsolins contain calcium-binding sites, which is manifested in calcium-controlled activities. Expression of two-domain gelsolins in mammalian cells enhanced actin filament disassembly on ionomycin-triggered calcium release. This functional demonstration, at the cellular level, provides evidence for a calcium-controlled Asgard actin cytoskeleton, indicating that the calcium-regulated actin cytoskeleton predates eukaryotes. In eukaryotes, dynamic bundled actin filaments are responsible for shaping filopodia and microvilli. By correlation, we hypothesize that the formation of the protrusions observed from Lokiarchaeota cell bodies may involve the gelsolin-regulated actin structures

Facial diplegia: etiology, clinical manifestations, and diagnostic evaluation

ABSTRACT Objective Facial diplegia (FD) is a rare neurological manifestation with diverse causes. This article aims to systematically evaluate the etiology, diagnostic evaluation and treatment of FD. Method The study was performed retrospectively and included 17 patients with a diagnosis of FD. Results Patients were diagnosed with Guillain-Barré syndrome (GBS) (11), Bickerstaff’s brainstem encephalitis (1), neurosarcoidosis (1), non-Hodgkin’s Lymphoma (1), tuberculous meningitis (1) herpes simplex reactivation (1) and idiopathic (1). In addition, two patients had developed FD during pregnancy. Conclusion Facial diplegia is an ominous symptom with widely varying causes that requires careful investigation

Structure and dynamics of Odinarchaeota tubulin and the implications for eukaryotic microtubule evolution

International audienceTubulins are critical for the internal organization of eukaryotic cells, and understanding their emergence is an important question in eukaryogenesis. Asgard archaea are the closest known prokaryotic relatives to eukaryotes. Here, we elucidated the apo and nucleotide-bound x-ray structures of an Asgard tubulin from hydrothermal living Odinarchaeota (OdinTubulin). The guanosine 5′-triphosphate (GTP)–bound structure resembles a microtubule protofilament, with GTP bound between subunits, coordinating the “+” end subunit through a network of water molecules and unexpectedly by two cations. A water molecule is located suitable for GTP hydrolysis. Time course crystallography and electron microscopy revealed conformational changes on GTP hydrolysis. OdinTubulin forms tubules at high temperatures, with short curved protofilaments coiling around the tubule circumference, more similar to FtsZ, rather than running parallel to its length, as in microtubules. Thus, OdinTubulin represents an evolutionary stage intermediate between prokaryotic FtsZ and eukaryotic microtubule-forming tubulins