Double Filaments Research Articles

Role of the antiparallel double-stranded filament form of FtsA in activating the Escherichia coli divisome.

The actin-like FtsA protein is essential for function of the cell division machinery, or divisome, in many bacteria including Escherichia coli. Previous in vitro studies demonstrated that purified wild-type FtsA assembles into closed mini-rings on lipid membranes, but oligomeric variants of FtsA such as FtsAR286W and FtsAG50E can bypass certain divisome defects and form arc and double-stranded (DS) oligomeric states, respectively, which may reflect conversion of an inactive to an active form of FtsA. However, it remains unproven which oligomeric forms of FtsA are responsible for assembling and activating the divisome. Here, we used an in vivo crosslinking assay for FtsA DS filaments to show that they largely depend on proper divisome assembly and are prevalent at later stages of cell division. We also used a previously reported variant that fails to assemble DS filaments, FtsAM96E R153D, to investigate the roles of FtsA oligomeric states in divisome assembly and activation. We show that FtsAM96E R153D cannot form DS filaments in vivo, fails to replace native FtsA, and confers a dominant negative phenotype, underscoring the importance of the DS filament stage for FtsA function. Surprisingly, however, activation of the divisome through the ftsL* or ftsW* superfission alleles suppressed the dominant negative phenotype and rescued the functionality of FtsAM96E R153D. Our results suggest that FtsA DS filaments are needed for divisome activation once it is assembled, but they are not essential for divisome assembly or guiding septum synthesis.IMPORTANCECell division is fundamental for cellular duplication. In simple cells like Escherichia coli bacteria, the actin homolog FtsA is essential for cell division and assembles into a variety of protein filaments at the cytoplasmic membrane. These filaments not only help tether polymers of the tubulin-like FtsZ to the membrane at early stages of cell division but also play crucial roles in recruiting other cell division proteins to a complex called the divisome. Once assembled, the E. coli divisome subsequently activates synthesis of the division septum that splits the cell in two. One recently discovered oligomeric conformation of FtsA is an antiparallel double-stranded filament. Using a combination of in vivo crosslinking and genetics, we provide evidence suggesting that these FtsA double filaments have a crucial role in activating the septum synthesis enzymes.

A method to produce periodic varied structural composite yarn by forced migrations of convergence point between polymer filaments and the staple strand

A novel composite yarn with a novel structure is prepared by the three-strand conversion method of double polymer filaments to staple fiber, that is, double polymer filaments to staple fiber repeated migration movement. The geometric principle of different relative positions of filament to helix structure change is analyzed theoretically, and the effect of filament position on the spinning-triangle of composite yarn is demonstrated. Furthermore, the structure of six different composite yarns is modeled and compared with each other, which shows the influence of different filament helical structures on the performance of spinning composite yarns. The repeated migration of filaments is observed using the online observation technique in the dynamic rotating triangle region. In addition, changes in yarn structure are caused by different geometric configurations. The results show that the stem, strength, and hair of six new composite yarns with different helical structures are slightly weakened. In summary, the three-strand conversion method is an effective new method to control the structure of filament and staple fibers.

Viscoelastic tensile model of core/wrapped composite yarn with double filament

Viscoelastic models are typically employed to investigate the mechanical properties of yarn. In this study, a viscoelastic model was developed to predict the tensile stress–strain relationship of a core/wrapped composite yarn with double filaments. The tensile properties of the yarn were tested, and various stages of the tensile curve were analyzed. Moreover, based on the tensile fracture characteristics, a five-element nonlinear viscoelastic model comprising Kelvin element, Maxwell element, and linear springs was established. Furthermore, the tensile properties of the composite yarns were simulated and calculated using the developed model. Additionally, the stress–strain relationship was fitted using a polynomial on the basis of the established model. The results reveal that the tensile fracture curve of the composite yarn comprises three stages. They are a small strain linear stage, a large strain stage, and a strength fluctuation stage. The viscoelastic tensile model can decently explain the three-stage stress–strain characteristics of the composite yarn tensile curve. The theoretical results were consistent with the experimental results, and the correlation coefficient was greater than 0.999. Based on the results, the proposed model can be employed with high accuracy to predict the tensile properties of a core/wrapped composite yarn with a double filament.

High Energy Release-high Retraction Smart Polymer Fibers used in Artificial Muscle Fabrication

Shape-memory polymers (SMPs) could remember their original shape and then, return to their initial shape upon stimulus. So far, quantities such as fixity and recovery ratio of SMPs have been broadly reported. Nevertheless, one main issue is the existence and use of an appropriate approach to quantitatively estimate the SMPs released energy. In addition, it is hypothesized that the elastic behavior of SMPs plays an underlying role when SMP fibers need to exhibit high-tension and high-elongation capacity as required in synthetic muscles. Here, we present, for first time, SMP trinary bulk and filament systems of acrylonitrile butadiene styrene (ABS)/thermoplastic polyurethane (TPU)/ethylene vinyl acetate (EVA) fabricated under calendering intense shear mixing and hot pressing. A digital blocked force load cell was used to record specimens energy released. The results exhibited high retraction from the specimens secondary shape correlated to the tensile behavior. It was shown the blended system of 50, 25 and 25% of TPU, ABS and EVA, respectively, resulted in ~640% and 3900% increase in the elastic modulus and energy release compared to EVA/TPU systems. The double SMP filaments led to a 50% increase in the energy release compared to single fibers. Nevertheless, the blended binary specimens with the TPU/ABS ratio of 50/50 exhibited 600% increase in tensile strength. It was confirmed the elastic modulus, number of fibers and elongation at break govern the SMP stored. The findings of the research lightened a new class of SMPs to be used as fiber-based artificial muscles and orthodontic products.

Femtosecond filamentation in water studied by the interference of supercontinuum

Supercontinuum (SC) generation is a typical nonlinear phenomenon that occurs during femtosecond filamentation in transparent media. The interference of the SC induced by femtosecond filament in water is explored by using interferometry with the aid of a Mach–Zehnder interferometer (MZI). In the low pulse energy case (single filament is formed), the MZI is used to precisely determine the actual value of filamentation threshold P th. It is found that the value of P th is much higher than that of the critical power for self-focusing P cr. In the higher pulse energy case, by blocking one arm of the MZI, self-interference resulted from SC emitted by femtosecond filaments is studied. By analyzing the interference patterns, we can acquire information on femtosecond filament, such as filament number and filament spacing. The energy range for the generation of single, double and triple filaments in water is also determined. Diffraction effect will distort and even mask the interference patterns, and to eliminate its influence on the results, the spectral signals at shorter wavelength are selected. This work provides an effective approach to study the complex femtosecond filamentation process intuitively and conveniently.

Bacterial divisome protein FtsA forms curved antiparallel double filaments when binding to FtsN.

During bacterial cell division, filaments of tubulin-like FtsZ form the Z-ring, which is the cytoplasmic scaffold for divisome assembly. In Escherichia coli, the actin homologue FtsA anchors the Z-ring to the membrane and recruits divisome components, including bitopic FtsN. FtsN regulates the periplasmic peptidoglycan synthase FtsWI. To characterize how FtsA regulates FtsN, we applied electron microscopy to show that E. coli FtsA forms antiparallel double filaments on lipid monolayers when bound to the cytoplasmic tail of FtsN. Using X-ray crystallography, we demonstrate that Vibrio maritimus FtsA crystallizes as an equivalent double filament. We identified an FtsA-FtsN interaction site in the IA-IC interdomain cleft of FtsA using X-ray crystallography and confirmed that FtsA forms double filaments in vivo by site-specific cysteine cross-linking. FtsA-FtsN double filaments reconstituted in or on liposomes prefer negative Gaussian curvature, like those of MreB, the actin-like protein of the elongasome. We propose that curved antiparallel FtsA double filaments together with treadmilling FtsZ filaments organize septal peptidoglycan synthesis in the division plane.

Immunosuppression and apoptosis activation mediated by p53-Bcl2/Bax signaling pathway -The potential mechanism of goldfish (Carassius auratus Linnaeus) gill disease caused by Myxobolus ampullicapsulatus.

Myxobolus, a major harmful type of myxospora, is one of the main parasitic pathogens of freshwater fish. Once myxoboliosis occurs, treatment can be extremely difficult. Therefore, clear understandings of the etiology of myxoboliosis and its pathological mechanism are keys for prevention and control. Here, histology, transmission electron microscopy, transcriptome study, tunel assay, and immunohistochemistry were carried out, revealing the morphology, pathological effects as well as host response mechanism of goldfish gill to Myxobolus ampullicapsulatus. Histological studies showed that the mature spores of Myxobolus ampullicapsulatus were composed of three parts, the spore shell, sporoplasm and bottle shaped polar capsule containing double S-shaped polar filaments. Transcriptome analysis revealed that Myxobolus ampullicapsulatus -infected (Myx) goldfish gills were characterized by apoptosis activation mediated by “p53 signaling pathway” with significantly up-regulated apoptosis-related differential genes dominated by p53-Bcl2/Bax signaling pathway. In addition, tunel assay revealed severe gill apoptosis in the Myx group. Transcriptome analysis also revealed that Myx group showed changes in immune response and significantly down-regulated immune-related differential genes. Beyond that, immunohistochemistry showed that there was no significant increase in the number of gill lymphocyte after parasite infection. These results suggest that the pathological mechanism of Myxobolus ampullicapsulatus infection on gills of goldfish may be related to apoptosis and immunosuppression. Subsequent qRT-PCR showed that apoptosis-related genes (Caspase3,Bad, Bax) and anti-inflammatory gene IL-10 were significantly increased, while immune-related pro-inflammatory genes (IL-1β, IL-8) were markedly down-regulated, further verifying the transcriptome results. Based on the above results, we concluded that p53-Bcl2/Bax related networks that dominant the expression of apoptosis genes were activated while immunity was suppressed in the gills of Myxobolus ampullicapsulatus infected goldfish. Our study is not only of benefit to enrich the taxonomy of Myxobolus but also clarifies its pathogenic mechanism, thus providing targets for prevention and control of myxoboliosis.

A Novel Z-Ring Associated Protein ZapA-Like Protein (PA5407) From Pseudomonas aeruginosa Promotes FtsZ to Form Double Filaments

Bacterial cell division is initiated by the assembly of the contraction ring (Z-ring), which consists of the self-assembled FtsZ protofilaments and dozens of other associate proteins. ZapA, a regulatory protein found in almost all bacteria, stabilizes FtsZ protofilaments to form bundles and enhances the Z-ring condensation. Here, we reported that another small protein from Pseudomonas aeruginosa, ZapA-Like protein (ZapAL; PA5407), is a new FtsZ associated protein. ZapAL exists in many Pseudomonas species and shares only 20% sequence identity to ZapA. ZapAL interacts with FtsZ and induces FtsZ to form long straight double filaments; in comparison, ZapA promotes long bundles with multiple FtsZ filaments. ZapAL has only a mild effect on GTPase activity of FtsZ, which is reduced by around 26% when 10 μM ZapAL is added in the solution. However, to study their assembly dynamics using light-scattering assay, we found that FtsZ-ZapAL double filament is stable and no depolymerization process is observed, which is different from ZapA. Further research found that ZapA and ZapL are likely to form heterodimers. The bundles formed by the mixture of FtsZ-ZapA-ZapAL will depolymerize after GTP is hydrolyzed. Consistent with ZapAL interaction with FtsZ in vitro, the expression of ZapAL-GFP was observed as a narrow band or spots in the middle of the cells, suggesting that it is a component of bacterial division machinery. Similar to ZapA, ZapAL is also not essential for bacterial cell division. Little changes were observed when zapAL gene was deleted, or overexpressed under normal conditions; however, overexpression of ZapAL caused zapA-deficient cells to grow approximately two times longer, showing a mild bacterial division defect. Although we still do not know the exact physiological roles of ZapAL, our results suggest that ZapAL is a novel Z-ring associate protein, which may work together with ZapA to stabilize the FtsZ protofilament and Z-ring structure.

Architectural Evaluation of 3D-Printed Buildings

3D-printed constructions express the capacity of automated technologies to elaborate buildings through additive manufacturing. These constructions require an architectural design according to their manufacturing conditions. This research reviews buildings that have been executed with these technologies in order to determine architectural characteristics. From an Internet search, a register was compiled of 112 cases of 3D-printed constructions around the globe. They include some 10,000 m2 of built surface area and were mainly erected in the last five years. The review shows that cases were built by approximately thirty executors, mostly entrepreneurs’ companies who have made different buildings as single-family dwellings constructed with gantries or robotic arms, either on-site or in factories. Most of the components printed are walls, with single or double filaments with interior voids. In some cases, they integrate reinforcements and openings. The geometry of the 3D-printed constructions varies between orthogonal layouts that replicate existing buildings and spherical shapes that reflect printing capabilities. Many of the cases are a combination of these characteristics, mixing a technological adaptation to existing architecture and new operative conditions. This review reveals the emerging development of this construction system with the progressive consolidation of some architectural attributes.

Filamentary damage of fused silica irradiated by a 532 nm nanosecond laser

In this study, we use a time-resolved pump-probe shadowgraph technique to investigate the evolution characteristics of filamentary damage in bulk fused silica induced by a nanosecond pulse at 532 nm. The pump laser focuses on the front surface of sample and filamentary damage appears independently in the middle of sample. The whole damage process can be divided into single filament (SF), double filaments (DFs), and long filament (LF) successively. At the same time, the improved moving focus model is proposed by taking into account the temporal shape of the laser pulse and the laser is blocked and reflected by plasma at the critical density. It is in good agreement with the experimental result of filamentary damage process and helps to explore the mechanism of laser-induced filamentary damage in nanosecond regime.

Optimisation of 186Os/188Os Measurements by N‐TIMS Using Amplifiers Equipped with 1013 Ω Resistors

Analysis of 186Os/188Os variations in most natural samples is hampered by their low Os mass fractions (usually < 100 pg g−1), requiring Os separation from up to a kilogram of powder to obtain high precision measurement results. Recently developed amplifiers equipped with 1013 Ω feedback resistors improve the signal to noise ratio by an order of magnitude, allowing acquisition of high precision data (2SE < 50 ppm) from 186Os16O3− ion beams of ~ 40 fA and thus analysis of less material (~ 10–500 g). Factors controlling the uncertainty of 186Os/188Os N‐TIMS measurement results, using a mixed array of 1013 and 1011 Ω amplifiers, include isobaric interferences, baseline variability, gain calibration, decay time corrections and oxygen isotope composition variations. Isobaric interferences present the greatest challenges, but these can be largely overcome by using double Pt filaments and periodically monitoring interferences during each measurement. The optimised procedures produce 186Os/188Os ratios with precisions close to the theoretical limits imposed by counting statistics and Johnson–Nyquist electronic noise for ion beams of 10–70 fA. The intermediate measurement precision (85 ppm; 2s) approaches theoretically expected values (~ 50–80 ppm). This precision is sufficient to allow 186Os/188Os analyses of crustal rocks, for which relatively large variations are expected.

Investigating the Influence of Architecture and Material Composition of 3D Printed Anatomical Design Scaffolds for Large Bone Defects.

There is a significant unmet clinical need to prevent amputations due to large bone loss injuries. We are addressing this problem by developing a novel, cost-effective osseointegrated prosthetic solution based on the use of modular pieces, bone bricks, made with biocompatible and biodegradable materials that fit together in a Lego-like way to form the prosthesis. This paper investigates the anatomical designed bone bricks with different architectures, pore size gradients, and material compositions. Polymer and polymer-composite 3D printed bone bricks are extensively morphological, mechanical, and biological characterized. Composite bone bricks were produced by mixing polycaprolactone (PCL) with different levels of hydroxyapatite (HA) and β-tri-calcium phosphate (TCP). Results allowed to establish a correlation between bone bricks architecture and material composition and bone bricks performance. Reinforced bone bricks showed improved mechanical and biological results. Best mechanical properties were obtained with PCL/TCP bone bricks with 38 double zig-zag filaments and 14 spiral-like pattern filaments, while the best biological results were obtained with PCL/HA bone bricks based on 25 double zig-zag filaments and 14 spiral-like pattern filaments.

3D-Printed Composite Bone Bricks For Large Bone Tissue Applications

This study investigates the use of low cost, customizable, biodegradable, polymer-ceramic composite porous structures (bone bricks) for large bone tissue regeneration. Different ceramic materials (hydroxyapatite (HA), β-tri-calcium phosphate (TCP) and Bioglass (45S5) were mixed with poly-ε-caprolactone (PCL). Bone bricks with different material compositions were produced using an extrusion-based additive manufacturing system. Produced bone bricks were morphologically and mechanically assessed. Results allowed to establish a correlation between scaffolds architecture and material composition and scaffolds performance. Reinforced scaffolds showed improved mechanical properties. Best mechanical properties were obtained with PCL/TCP bone bricks and topologies based on 38 double zig zag filaments and 14 spirals.

Group-Sparsity-Based Super-Resolution Dipole Orientation Mapping.

The dipole orientation of fluorophores could be resolved by fluorescence polarization microscopy (FPM), which in turn reveals structural specificity for the labeled organelles. Conventional FPM can detect only the averaged fluorescence anisotropy collected from dipoles within the diffraction-limited volume. Super-resolution dipole orientation mapping (SDOM) method, which applies sparse deconvolution and least square estimation to fluorescence polarization modulation data, achieves the dipole orientation measurement within a sub-diffraction focal area. However, during SDOM analysis, some pixels with fluorescence signal are not resolved with orientation for relatively small adjusted R2. Here we report group-sparsity-based SDOM (GS-SDOM), which utilizes the relevance of modulation sequences to effectively improve the SDOM reconstruction model. More credible resolved dipole orientations with higher adjusted R2 can be mapped and false positive estimation for local dipole orientation is vitally corrected. In addition to achieving the same spatial super-resolution as SDOM does, GS-SDOM accesses more morphological information with more credible orientations and more accurate local dipole distribution estimation. During the GS-SDOM analysis of actin filaments in mammalian kidney cells, the dipole orientation of fluorescence is detected always parallel to the direction of the actin filaments. Also with dipole orientation information extracted by GS-SDOM, the reconstructed visual circle from intensity dimension is discerned as jointed by double close filaments and 3-dimensional co-localization is accomplished in the intersection of actin filaments.

Gain-of-function variants of FtsA form diverse oligomeric structures on lipids and enhance FtsZ protofilament bundling.

Escherichia coli requires FtsZ, FtsA and ZipA proteins for early stages of cell division, the latter two tethering FtsZ polymers to the cytoplasmic membrane. Hypermorphic mutants of FtsA such as FtsA* (R286W) map to the FtsA self-interaction interface and can bypass the need for ZipA. Purified FtsA forms closed minirings on lipid monolayers that antagonize bundling of FtsZ protofilaments, whereas FtsA* forms smaller oligomeric arcs that enable bundling. Here, we examined three additional FtsA*-like mutant proteins for their ability to form oligomers on lipid monolayers and bundle FtsZ. Surprisingly, all three formed distinct structures ranging from mostly arcs (T249M), a mixture of minirings, arcs and straight filaments (Y139D) or short straight double filaments (G50E). All three could form filament sheets at higher concentrations with added ATP. Despite forming these diverse structures, all three mutant proteins acted like FtsA* to enable FtsZ protofilament bundling on lipid monolayers. Synthesis of the FtsA*-like proteins in vivo suppressed the toxic effects of a bundling-defective FtsZ, exacerbated effects of a hyper-bundled FtsZ, and rescued some thermosensitive cell division alleles. Together, the data suggest that conversion of FtsA minirings into any type of non-miniring oligomer can promote progression of cytokinesis through FtsZ bundling and other mechanisms.

Cryo-EM reconstruction of AlfA from Bacillus subtilis reveals the structure of a simplified actin-like filament at 3.4-Å resolution

Low copy-number plasmid pLS32 of Bacillus subtilis subsp. natto contains a partitioning system that ensures segregation of plasmid copies during cell division. The partitioning locus comprises actin-like protein AlfA, adaptor protein AlfB, and the centromeric sequence parN Similar to the ParMRC partitioning system from Escherichia coli plasmid R1, AlfA filaments form actin-like double helical filaments that arrange into an antiparallel bipolar spindle, which attaches its growing ends to sister plasmids through interactions with AlfB and parN Because, compared with ParM and other actin-like proteins, AlfA is highly diverged in sequence, we determined the atomic structure of nonbundling AlfA filaments to 3.4-Å resolution by cryo-EM. The structure reveals how the deletion of subdomain IIB of the canonical actin fold has been accommodated by unique longitudinal and lateral contacts, while still enabling formation of left-handed, double helical, polar and staggered filaments that are architecturally similar to ParM. Through cryo-EM reconstruction of bundling AlfA filaments, we obtained a pseudoatomic model of AlfA doublets: the assembly of two filaments. The filaments are antiparallel, as required by the segregation mechanism, and exactly antiphasic with near eightfold helical symmetry, to enable efficient doublet formation. The structure of AlfA filaments and doublets shows, in atomic detail, how deletion of an entire domain of the actin fold is compensated by changes to all interfaces so that the required properties of polymerization, nucleotide hydrolysis, and antiparallel doublet formation are retained to fulfill the system's biological raison d'être.

Crenactin forms actin-like double helical filaments regulated by arcadin-2.

The similarity of eukaryotic actin to crenactin, a filament-forming protein from the crenarchaeon Pyrobaculum calidifontis supports the theory of a common origin of Crenarchaea and Eukaryotes. Monomeric structures of crenactin and actin are similar, although their filament architectures were suggested to be different. Here we report that crenactin forms bona fide double helical filaments that show exceptional similarity to eukaryotic F-actin. With cryo-electron microscopy and helical reconstruction we solved the structure of the crenactin filament to 3.8 Å resolution. When forming double filaments, the 'hydrophobic plug' loop in crenactin rearranges. Arcadin-2, also encoded by the arcade gene cluster, binds tightly with its C-terminus to the hydrophobic groove of crenactin. Binding is reminiscent of eukaryotic actin modulators such as cofilin and thymosin β4 and arcadin-2 is a depolymeriser of crenactin filaments. Our work further supports the theory of shared ancestry of Eukaryotes and Crenarchaea.

Pre-amyloid oligomers of the proteotoxic RepA-WH1 prionoid assemble at the bacterial nucleoid.

Upon binding to short specific dsDNA sequences in vitro, the N-terminal WH1 domain of the plasmid DNA replication initiator RepA assembles as amyloid fibres. These are bundles of single or double twisted tubular filaments in which distorted RepA-WH1 monomers are the building blocks. When expressed in Escherichia coli, RepA-WH1 triggers the first synthetic amyloid proteinopathy in bacteria, recapitulating some of the features of mammalian prion diseases: it is vertically transmissible, albeit non-infectious, showing up in at least two phenotypically distinct and interconvertible strains. Here we report B3h7, a monoclonal antibody specific for oligomers of RepA-WH1, but which does not recognize the mature amyloid fibres. Unlike a control polyclonal antibody generated against the soluble protein, B3h7 interferes in vitro with DNA-promoted or amyloid-seeded assembly of RepA-WH1 fibres, thus the targeted oligomers are on-pathway amyloidogenic intermediates. Immuno-electron microscopy with B3h7 on thin sections of E. coli cells expressing RepA-WH1 consistently labels the bacterial nucleoid, but not the large cytoplasmic aggregates of the protein. This observation points to the nucleoid as the place where oligomeric amyloid precursors of RepA-WH1 are generated, and suggests that, once nucleated by DNA, further growth must continue in the cytoplasm due to entropic exclusion.

Amyloidogenesis of Bacterial Prionoid RepA-WH1 Recapitulates Dimer to Monomer Transitions of RepA in DNA Replication Initiation

Most available structures of amyloids correspond to peptide fragments that self-assemble in extended cross β sheets. However, structures in which a whole protein domain acts as building block of an amyloid fiber are scarce, in spite of their relevance to understand amyloidogenesis. Here, we use electron microscopy (EM) and atomic force microscopy (AFM) to analyze the structure of amyloid filaments assembled by RepA-WH1, a winged-helix domain from a DNA replication initiator in bacterial plasmids. RepA-WH1 functions as a cytotoxic bacterial prionoid that recapitulates features of mammalian amyloid proteinopathies. RepA are dimers that monomerize at the origin to initiate replication, and we find that RepA-WH1 reproduces this transition to form amyloids. RepA-WH1 assembles double helical filaments by lateral association of a single-stranded precursor built by monomers. Double filaments then associate in mature fibers. The intracellular and cytotoxic RepA-WH1 aggregates might reproduce the hierarchical assembly of human amyloidogenic proteins.

Architecture and dynamic remodelling of the septin cytoskeleton during the cell cycle.

Septins perform diverse functions through the formation of filaments and higher-order structures. However, the exact architecture of septin structures remains unclear. In the budding yeast Saccharomyces cerevisiae, septins form an “hourglass” at the mother-bud neck before cytokinesis, which is converted into a “double ring” during cytokinesis. Here, using platinum-replica electron microscopy, we find that the early hourglass consists of septin double filaments oriented along the mother-bud axis. In the late hourglass, these double filaments are connected by periodic circumferential single filaments on the membrane-proximal side, and are associated with centrally located, circumferential, myosin-II thick filaments on the membrane-distal side. The double ring consists of exclusively circumferential septin filaments. Live-cell imaging studies indicate that the hourglass-to-double ring transition is accompanied by loss of septin subunits from the hourglass, and reorganization of the remaining subunits into the double ring. This work provides an unparalleled view of septin structures within cells and defines their remodeling dynamics during the cell cycle.