For construction of the bacterial flagellum, which is responsible for bacterial motility, the flagellar type III export apparatus utilizes both ATP and proton motive force across the cytoplasmic ...membrane and exports flagellar proteins from the cytoplasm to the distal end of the nascent structure. The export apparatus consists of a membrane-embedded export gate made of FlhA, FlhB, FliO, FliP, FliQ, and FliR and a water-soluble ATPase ring complex consisting of FliH, FliI, and FliJ. FlgN, FliS, and FliT act as substrate-specific chaperones that do not only protect their cognate substrates from degradation and aggregation in the cytoplasm but also efficiently transfer the substrates to the export apparatus. The ATPase ring complex facilitates the initial entry of the substrates into the narrow pore of the export gate. The export gate by itself is a proton-protein antiporter that uses the two components of proton motive force, the electric potential difference and the proton concentration difference, for different steps of the export process. A specific interaction of FlhA with FliJ located in the center of the ATPase ring complex allows the export gate to efficiently use proton motive force to drive protein export. The ATPase ring complex couples ATP binding and hydrolysis to its assembly–disassembly cycle for rapid and efficient protein export cycle. This article is part of a Special Issue entitled: Protein trafficking and secretion in bacteria. Guest Editors: Anastassios Economou and Ross Dalbey.
•Most flagellar proteins are exported by a specific export apparatus.•The export apparatus consists of an export gate and an ATPase complex.•The export apparatus coordinates protein export with flagellar assembly.•Flagellar chaperones efficiently transfer their substrates to the export apparatus.•The export apparatus uses both ATP and proton motive force for rapid protein export.
The bacterial flagellum is supramolecular motility machinery consisting of the basal body, the hook and the filament. Flagellar proteins are translocated across the cytoplasmic membrane via a type ...III protein export apparatus, diffuse down the central channel of the growing structure and assemble at the distal end. Flagellar assembly begins with the basal body, followed by the hook and finally the filament. The completion of hook assembly is the most important morphological checkpoint of the sequential flagellar assembly process. When the hook reaches its mature length of about 55 nm in Salmonella enterica, the type III protein export apparatus switches export specificity from proteins required for the structure and assembly of the hook to those responsible for filament assembly, thereby terminating hook assembly and initiating filament assembly. Three flagellar proteins, namely FliK, FlhB and FlhA, are responsible for this substrate specificity switching. Upon completion of the switching event, interactions among FlhA, the cytoplasmic ATPase complex and flagellar type III export chaperones establish the assembly order of the filament at the hook tip. Here, we describe our current understanding of a hierarchical protein export mechanism used in flagellar type III protein export.
Abstract The heart must continuously pump blood to supply the body with oxygen and nutrients. To maintain the high energy consumption required by this role, the heart is equipped with multiple ...complex biological systems that allow adaptation to changes of systemic demand. The processes of growth (hypertrophy), angiogenesis, and metabolic plasticity are critically involved in maintenance of cardiac homeostasis. Cardiac hypertrophy is classified as physiological when it is associated with normal cardiac function or as pathological when associated with cardiac dysfunction. Physiological hypertrophy of the heart occurs in response to normal growth of children or during pregnancy, as well as in athletes. In contrast, pathological hypertrophy is induced by factors such as prolonged and abnormal hemodynamic stress, due to hypertension, myocardial infarction etc. Pathological hypertrophy is associated with fibrosis, capillary rarefaction, increased production of pro-inflammatory cytokines, and cellular dysfunction (impairment of signaling, suppression of autophagy, and abnormal cardiomyocyte/non-cardiomyocyte interactions), as well as undesirable epigenetic changes, with these complex responses leading to maladaptive cardiac remodeling and heart failure. This review describes the key molecules and cellular responses involved in physiological/pathological cardiac hypertrophy.
The bacterial flagellum is a helical filamentous organelle responsible for motility. In bacterial species possessing flagella at the cell exterior, the long helical flagellar filament acts as a ...molecular screw to generate thrust. Meanwhile, the flagella of spirochetes reside within the periplasmic space and not only act as a cytoskeleton to determine the helicity of the cell body, but also rotate or undulate the helical cell body for propulsion. Despite structural diversity of the flagella among bacterial species, flagellated bacteria share a common rotary nanomachine, namely the flagellar motor, which is located at the base of the filament. The flagellar motor is composed of a rotor ring complex and multiple transmembrane stator units and converts the ion flux through an ion channel of each stator unit into the mechanical work required for motor rotation. Intracellular chemotactic signaling pathways regulate the direction of flagella-driven motility in response to changes in the environments, allowing bacteria to migrate towards more desirable environments for their survival. Recent experimental and theoretical studies have been deepening our understanding of the molecular mechanisms of the flagellar motor. In this review article, we describe the current understanding of the structure and dynamics of the bacterial flagellum.
Highlights • The rotor and stator of the flagellar motor are highly-dynamic structures. • The export components show dynamic interactions with their binding partners. • The assembly mechanism of the ...core structure of the motor is conserved. • There is structural diversity in the motor across bacterial species.
•Cardiac cells become senescent with aging.•Cellular senescence is mostly pathological in failing hearts.•Senolytics reverse aging phenotype in dysfunctional hearts.
Replicative capacity of somatic ...cells is limited. It indicates that aging also develops at the cellular level, and this is described as “cellular senescence”. Senescent cells become flattened, enlarged, and irreversibly lose capacity for proliferation. Lack of specific and conclusive markers for cellular senescence makes it difficult to comprehensively define and understand this biological process especially in vivo. Molecules including p53, p21, p16Ink4a, p38MAPK, and γH2AX, telomere attrition, enhanced signals for SA-β-gal, etc. are widely used to detect senescent cells, but these are indirect indicators of cellular senescence, and biological markers reflecting direct evidence need to be established. Genetic profiles are altered in senescent cells, letting these cells secrete pro-inflammatory molecules. Aging or age-related disorders including heart failure and atherosclerotic diseases link with an accumulation of cells undergoing cellular senescence in cardiovascular systems including heart and vessels. Senescent cells become pathogenic in most cases by mediating chronic sterile inflammation and tissue remodeling. A recent conceptual as well as technical breakthrough in this research area is “senolysis”, meaning the specific elimination of senescent cells. Genetic as well as pharmacological models with senolysis contributed to reverse aging phenotypes and ameliorated pathologies in age-related disorders without enhancing the risk of tumorigenesis, and opened a new avenue for aging research. Several compounds are identified as senolytics, and some are already tested in clinical settings. It was recently reported that senolysis reverses aging phenotype in cardiovascular disorders. Generating therapies targeting suppression or elimination of senescent cells would inhibit the progression of undesirable aspects of aging, and become promising therapies for cardiac diseases.
Abstract
The basal body of the bacterial flagellum is a rotary motor that consists of several rings (C, MS and LP) and a rod. The LP ring acts as a bushing supporting the distal rod for its rapid and ...stable rotation without much friction. Here, we use electron cryomicroscopy to describe the LP ring structure around the rod, at 3.5 Å resolution, from
Salmonella
Typhimurium. The structure shows 26-fold rotational symmetry and intricate intersubunit interactions of each subunit with up to six partners, which explains the structural stability. The inner surface is charged both positively and negatively. Positive charges on the P ring (the part of the LP ring that is embedded within the peptidoglycan layer) presumably play important roles in its initial assembly around the rod with a negatively charged surface.
Translocation of many soluble proteins across cell membranes occurs in an ATPase-driven manner. For construction of the bacterial flagellum responsible for motility, most of the components are ...exported by the flagellar protein export apparatus. The FliI ATPase is required for this export, and its ATPase activity is regulated by FliH; however, it is unclear how the chemical energy derived from ATP hydrolysis is used for the export process. Here we report that flagellar proteins of Salmonella enterica serovar Typhimurium are exported even in the absence of FliI. A fliH fliI double null mutant was weakly motile. Certain mutations in FlhA or FlhB, which form the core of the export gate, substantially improved protein export and motility of the double null mutant. Furthermore, proton motive force was essential for the export process. These results suggest that the FliH-FliI complex facilitates only the initial entry of export substrates into the gate, with the energy of ATP hydrolysis being used to disassemble and release the FliH-FliI complex from the protein about to be exported. The rest of the successive unfolding/translocation process of the substrates is driven by proton motive force.
Celotno besedilo
Dostopno za:
DOBA, IJS, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The bacterial flagellar motor is embedded within the cell envelop and rotates the long helical filament, which acts as a molecular screw to propel the bacterium. The flagellar motor comprises a rotor ...and a dozen stator units, converting ion flux through the stator unit into torque. However, the energy coupling mechanism has not been fully understood. Various methods for rotation measurement have been developed to understand the rotation mechanism of the flagellar motor, but the most preferred method in recent studies is a bead assay, which tracks the rotation of a micron to submicron bead attached to the partially sheared flagellar filament at high temporal and spatial resolutions. The bead assay allows us to assess the motor rotation over a wide range of external load, but the elasticity of the axial parts of the flagellum, such as the hook and filament, limits the spatiotemporal resolution. In this chapter, we describe a bead assay optimized for the analysis of the flagellar motor dynamics at near zero load.
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