Ring Complex Research Articles

Nonlinear optical properties of symmetrical and asymmetrical benzene derivatives with click chemistry modification

PurposeThis paper aims to prepare third-order nonlinear optical (NLO) organic materials with large nonlinear optimization value, high damage threshold and ultrafast response time.Design/methodology/approachA series of novel symmetric and asymmetric compounds possessing third-order NLO properties were synthesized using 1,3,5-tribromobenzene as the basis. The photophysical and electrochemical properties, as well as the click reactions, were characterized by means of UV–VIS–NIR absorption spectroscopy and cyclic voltammetry.FindingsThe donor–acceptor chromophores were inserted into compound, making the molecule to have a broader absorption in the near-infrared regions and a narrower optical and electrochemical band gap. It also formed an electron-delocalized organic system, which has larger effects on achieving a third-order NLO response. The third-order NLO phenomenon of benzene ring complexes was experimentally studied at 532 nm using Z-scan technology, and some compounds showed the expected NLO properties.Originality/valueThe click products exhibit more NLO phenomena by performing different click combinations to the side groups, opening new perspectives on using the system in a variety of photoelectric applications.

Tectonic evolution of the neoproterozoic tusham ring complex, Northwestern India: Constraints from geochemistry and zircon U Pb geochronology, and implications for Rodinia supercontinent history

Tectonic evolution of the neoproterozoic tusham ring complex, Northwestern India: Constraints from geochemistry and zircon U Pb geochronology, and implications for Rodinia supercontinent history

The Role of TRiC-enhanced Actin Folding in Leber Congenital Amaurosis.

Mutations in TCP-1 ring complex (TRiC) have been associated with Leber Congenital Amaurosis (LCA). TRiC is involved in protein folding and has 8 essential subunits including CCT5. Herein, we studied the retina of TRiC mutant zebrafish to evaluate the possible role of impaired actin and tubulin folding in LCA. The cct5 retina was histologically studied using Toluidine Blue staining as well as TUNEL, BrdU-labeling, and Phalloidin assays. Retinal organisation was assessed by quantification of the cellularity utilising DAPI. Laminar organization of cct5 retinas was intact. Enhanced apoptosis throughout the cct5 retina was not compensated by higher proliferation rates, leaving the cct5 retina smaller in size. Quantification of retinal layer cellularity demonstrated that specifically the numbers of the amacrine and the retinal ganglion cells were depleted, suggesting that the cct5 retina was not uniformly affected by the reduced actin folding. Whereas the current literature suggests that LCA is predominantly affecting retinal photoreceptor cells and the retinal pigment epithelium, cct5 analyses demonstrated the important role of folding of actin by TRiC, suggesting that cct5 is a useful tool to specifically analyze the role of F-actin filaments in the context of LCA.

A structural mechano-chemical model of microtubule nucleation.

Microtubules (MTs) are the backbone of cytoskeleton and crucial for many essential processes, such as mitosis, morphogenesis and migration. A critical step in the assembly and regulation of cellular MT network is nucleation. There are two types of templates for MT nucleation in cells: existing MT and γ-tubulin ring complex. Both types of nucleations have been reconstituted in vitro, but their mechanisms remain poorly understood. Here, we developed a physically rigorous structure-based mechano-chemical model that provides an unified mechanism for both types of nucleations. For nucleation from seed MTs, the key observations are: 1) the critical tubulin concentration for nucleation is much higher than that for growth, 2) the concentration-dependent time lags. Our model shows that the bottleneck for nucleation is frequent catastrophes (i.e., the switching from MT growth to shortening) that keep removing nascent MTs from the seed, which is reduced as tubulin concentration increases. The observed time lag is due to repeated assembly and removal of nascent MTs, which delay nucleation. For nucleation from γ-tubulin ring, the most intriguing question is the effects of ring closure, a complex-wide conformational change required for the ring complex to match the tube-like geometry of MT. Our model shows that ring closure helps tubulins to first form a sheet, which is then closed into tube with the help of the ring closure process. Because catastrophe cannot develop while sheet is open, MTs more likely grow longer, increasing the chance of nucleation. Thus, ring closure allows more regulated and efficient MT nucleation.

Breaking down the HAUS: Molecular insights into branching microtubule nucleation.

Accurate segregation of chromosomes during mitosis depends on the correct assembly of the mitotic spindle, a bipolar structure composed mainly of microtubules. The augmin complex, or homologous to augmin subunits (HAUS) complex, is an eight-subunit protein complex required for building robust mitotic spindles in metazoa. Augmin increases microtubule density within the spindle by recruiting the γ-tubulin ring complex (γ-TuRC) to pre-existing microtubules and nucleating branching microtubules in a manner that maintains spindle polarity. Here, we elucidate the molecular architecture of augmin by single particle cryo-electron microscopy (cryo-EM), computational methods, and crosslinking mass spectrometry (CLMS). Augmin's highly flexible structure contains a V-shaped head and a filamentous tail, with the head existing in either extended or contracted conformational states. Our work highlights how cryo-EM, complemented by computational advances and CLMS, can elucidate the structure of a challenging protein complex. We demonstrate how this approach has provided novel insights into the function of augmin in mediating microtubule branching nucleation.

Mineralogical and Geochemical Evidence of Paragenetic Unity of Igneous Silicate and Carbonatite Rocks of the Tomtor Massif in the North-East of the Siberian Platform

The Tomtor massif is a polychronous ring zonal complex of alkaline ultramafic and carbonatite rocks containing unique Nb and REE deposits. Mineralogical and geochemical studies of minerals from different types of silicate rocks and carbonatites of the Tomtor massif were performed. For excluding traces of the interaction between silicate and carbonatite melts, we limited ourselves to the study of independent small secant bodies located in the immediate vicinity of the massif itself. The presence of through mineral series in various silicate igneous rocks and carbonatite ores of high-titanium chromium spinels, rare-metal, ore and other exotic phases with similar compositional trends was defined. Such studies will help reveal the mineralogical criteria for the genetic relationship between silicate melts and associated carbonatite derivatives, which can form rich rare elements mineralization. Also, such studies help to improve the petrochemical and mineralogical criteria for dividing potentially diamond-bearing magmatites (typical kimberlites) from non-diamond-bearing kimberlites, alpicrites and other non-diamond-bearing rocks convergent to kimberlites, which are formed under different physicochemical conditions. The existence of polychronous complex ore–magmatic ring complexes, such as the Tomtor massif, indicates the existence of large deep intraplate magma-generating chambers in the lithospheric mantle.

Proper phosphorylation of septin 12 regulates septin 4 and soluble adenylyl cyclase expression to induce sperm capacitation.

Septin-based ring complexes maintain the sperm annulus. Defective annular structures are observed in the sperm of Sept12- and Sept4-null mice. In addition, sperm capacitation, a process required for proper fertilization, is inhibited in Sept4-null mice, implying that the sperm annulus might play a role in controlling sperm capacitation. Hence, we analyzed sperm capacitation of sperm obtained from SEPT12 Ser196 phosphomimetic (S196E), phosphorylation-deficient (S196A), and SEPT4-depleted mutant mice. Capacitation was reduced in the sperm of both the Sept12 S196E- and Sept12 S196A-knock-in mice. The protein levels of septins, namely, SEPT4 and SEPT12, were upregulated, and these proteins were concentrated in the sperm annulus during capacitation. Importantly, the expression of soluble adenylyl cyclase (sAC), a key enzyme that initiates capacitation, was upregulated, and sAC was recruited to the sperm annulus following capacitation stimulation. We further found that SEPT12, SEPT4, and sAC formed a complex and colocalized to the sperm annulus. Additionally, sAC expression was reduced and disappeared in the annulus of the SEPT12 S196E- and S196A-mutant mouse sperm. In the sperm of the SEPT4-knockout mice, sAC did not localize to the annulus. Thus, our data demonstrate that SEPT12 phosphorylation status and SEPT4 activity jointly regulate sAC protein levels and annular localization to induce sperm capacitation.

Microtubule nucleation and γTuRC centrosome localization in interphase cells require ch-TOG

Organization of microtubule arrays requires spatio-temporal regulation of the microtubule nucleator γ-tubulin ring complex (γTuRC) at microtubule organizing centers (MTOCs). MTOC-localized adapter proteins are thought to recruit and activate γTuRC, but the molecular underpinnings remain obscure. Here we show that at interphase centrosomes, rather than adapters, the microtubule polymerase ch-TOG (also named chTOG or CKAP5) ultimately controls γTuRC recruitment and activation. ch-TOG co-assembles with γTuRC to stimulate nucleation around centrioles. In the absence of ch-TOG, γTuRC fails to localize to these sites, but not the centriole lumen. However, whereas some ch-TOG is stably bound at subdistal appendages, it only transiently associates with PCM. ch-TOG’s dynamic behavior requires its tubulin-binding TOG domains and a C-terminal region involved in localization. In addition, ch-TOG also promotes nucleation from the Golgi. Thus, at interphase centrosomes stimulation of nucleation and γTuRC attachment are mechanistically coupled through transient recruitment of ch-TOG, and ch-TOG’s nucleation-promoting activity is not restricted to centrosomes.

A nucleotide binding-independent role for γ-tubulin in microtubule capping and cell division.

The γ-tubulin ring complex (γ-TuRC) has essential roles in centrosomal and non-centrosomal microtubule organization during vertebrate mitosis. While there have been important advances in understanding γ-TuRC-dependent microtubule nucleation, γ-TuRC capping of microtubule minus-ends remains poorly characterized. Here, we utilized biochemical reconstitutions and cellular assays to characterize the human γ-TuRC's capping activity. Single filament assays showed that the γ-TuRC remained associated with a nucleated microtubule for tens of minutes. In contrast, caps at dynamic microtubule minus-ends displayed lifetimes of ∼1 min. Reconstituted γ-TuRCs with nucleotide-binding deficient γ-tubulin (γ-tubulinΔGTP) formed ring-shaped complexes that did not nucleate microtubules but capped microtubule minus-ends with lifetimes similar to those measured for wild-type complexes. In dividing cells, microtubule regrowth assays revealed that while knockdown of γ-tubulin suppressed non-centrosomal microtubule formation, add-back of γ-tubulinΔGTP could substantially restore this process. Our results suggest that γ-TuRC capping is a nucleotide-binding-independent activity that plays a role in non-centrosomal microtubule organization during cell division.

Chaperonin TRiC/CCT Participates in Mammarenavirus Multiplication in Human Cells via Interaction with the Viral Nucleoprotein.

The eukaryotic chaperonin containing tailless complex polypeptide 1 ring complex (CCT, also known as TCP-1 Ring Complex, TRiC/CCT) participates in the folding of 5% to 10% of the cellular proteome and has been involved in the life cycle of several viruses, including dengue, Zika, and influenza viruses, but the mechanisms by which the TRiC/CCT complex contributes to virus multiplication remain poorly understood. Here, we document that the nucleoprotein (NP) of the mammarenavirus lymphocytic choriomeningitis virus (LCMV) is a substrate of the human TRiC/CCT complex, and that pharmacological inhibition of TRiC/CCT complex function, or RNAi-mediated knockdown of TRiC/CCT complex subunits, inhibited LCMV multiplication in human cells. We obtained evidence that the TRiC/CCT complex is required for the production of NP-containing virus-like particles (VLPs), and the activity of the virus ribonucleoprotein (vRNP) responsible for directing replication and transcription of the viral genome. Pharmacological inhibition of the TRIC/CCT complex also restricted multiplication of the live-attenuated vaccine candidates Candid#1 and ML29 of the hemorrhagic fever causing Junin (JUNV) and Lassa (LASV) mammarenaviruses, respectively. Our findings indicate that the TRiC/CCT complex is required for mammarenavirus multiplication and is an attractive candidate for the development of host directed antivirals against human-pathogenic mammarenaviruses. IMPORTANCE Host-directed antivirals have gained great interest as an antiviral strategy to counteract the rapid emergence of drug-resistant viruses. The chaperonin TRiC/CCT complex has been involved in the life cycle of several viruses, including dengue, Zika, and influenza viruses. Here, we have provided evidence that the chaperonin TRiC/CCT complex participates in mammarenavirus infection via its interaction with the viral NP. Importantly, pharmacological inhibition of TRiC/CCT function significantly inhibited multiplication of LCMV and the distantly related mammarenavirus JUNV in human cells. Our findings support that the TRiC/CCT complex is required for multiplication of mammarenaviruses and that the TRiC/CCT complex is an attractive host target for the development of antivirals against human-pathogenic mammarenaviruses.

Effects of major and trace elements from the El Kahfa ring complex on fish: Geological, physicochemical, and biological approaches

The alkaline rocks are known for enriching rare lithophilic elements, including lithium, uranium, and tin, which negatively impact aquatic life. This study offers an intensive investigation of the influence of alkaline rocks on Nile Tilapia (Oreochromis niloticus). The variation in blood profile, the induction of antioxidant enzymes, morphological erythrocyte, and histological structure have been conducted for the fish after 15 days of exposure to alkaline rocks powder with a dose of 100 μg/L. As a result, there was a pronounced decrease in blood profiles, such as platelets and white blood cell counts. There was a failure in the liver and kidney functions. Moreover, it shows an increase in superoxide dismutase (SOD) and catalase (CAT) activities as antioxidant biomarkers. Also, exposure to alkaline rocks induced DNA mutation and erythrocyte distortion. We concluded that the bulk alkaline rocks induced changes in the hemato-biochemical and antioxidant parameters of Nile tilapia. Additionally, exposure to bulk alkaline rock compounds also caused poikilocytosis and nuclear abnormalities of RBCs. This draws our attention to the seriousness of climatic changes, the erosion of rocks, and their access to water.

“Proteotranscriptomic analysis of advanced colorectal cancer patient derived organoids for drug sensitivity prediction”

BackgroundPatient-derived organoids (PDOs) from advanced colorectal cancer (CRC) patients could be a key platform to predict drug response and discover new biomarkers. We aimed to integrate PDO drug response with multi-omics characterization beyond genomics.MethodsWe generated 29 PDO lines from 22 advanced CRC patients and provided a morphologic, genomic, and transcriptomic characterization. We performed drug sensitivity assays with a panel of both standard and non-standard agents in five long-term cultures, and integrated drug response with a baseline proteomic and transcriptomic characterization by SWATH-MS and RNA-seq analysis, respectively.ResultsPDOs were successfully generated from heavily pre-treated patients, including a paired model of advanced MSI high CRC deriving from pre- and post-chemotherapy liver metastasis. Our PDOs faithfully reproduced genomic and phenotypic features of original tissue. Drug panel testing identified differential response among PDOs, particularly to oxaliplatin and palbociclib. Proteotranscriptomic analyses revealed that oxaliplatin non-responder PDOs present enrichment of the t-RNA aminoacylation process and showed a shift towards oxidative phosphorylation pathway dependence, while an exceptional response to palbociclib was detected in a PDO with activation of MYC and enrichment of chaperonin T-complex protein Ring Complex (TRiC), involved in proteome integrity. Proteotranscriptomic data fusion confirmed these results within a highly integrated network of functional processes involved in differential response to drugs.ConclusionsOur strategy of integrating PDOs drug sensitivity with SWATH-mass spectrometry and RNA-seq allowed us to identify different baseline proteins and gene expression profiles with the potential to predict treatment response/resistance and to help in the development of effective and personalized cancer therapeutics.

A quantitative map of nuclear pore assembly reveals two distinct mechanisms

Understanding how the nuclear pore complex (NPC) is assembled is of fundamental importance to grasp the mechanisms behind its essential function and understand its role during the evolution of eukaryotes1–4. There are at least two NPC assembly pathways—one during the exit from mitosis and one during nuclear growth in interphase—but we currently lack a quantitative map of these events. Here we use fluorescence correlation spectroscopy calibrated live imaging of endogenously fluorescently tagged nucleoporins to map the changes in the composition and stoichiometry of seven major modules of the human NPC during its assembly in single dividing cells. This systematic quantitative map reveals that the two assembly pathways have distinct molecular mechanisms, in which the order of addition of two large structural components, the central ring complex and nuclear filaments are inverted. The dynamic stoichiometry data was integrated to create a spatiotemporal model of the NPC assembly pathway and predict the structures of postmitotic NPC assembly intermediates.

Understanding electrostatics and covalency effects in highly anisotropic organometallic sandwich dysprosium complexes [Dy(CmRm)2] (where R = H, SiH3, CH3 and m = 4 to 9): a computational perspective.

In this article, we have thoroughly studied the electronic structure and 4f-ligand covalency of six mononuclear dysprosium organometallic sandwich complexes [Dy(CmRm)2]n+/- (where R = H, SiH3, CH3; m = 4 to 9; n = 1, 3) using both the scalar relativistic density functional and complete active space self-consistent field (CASSCF) and N-electron valence perturbation theory (NEVPT2) method to shed light on the ligand field effects in fine-tuning the magnetic anisotropy of these complexes. Energy decomposition analysis (EDA) and ab initio-based ligand field theory AILFT calculations predict the sizable 4f-ligand covalency in all these complexes. The analysis of CASSCF/NEVPT2 computed spin-Hamiltonian (SH) parameters indicates the stabilization of mJ |±15/2〉 for [Dy(C4(SiH3)4)2]- (1), [Dy(C5(CH3)5)2]+ (2) and [Dy(C6H6)2]3+ (3) complexes with the Ucal value of 1867.5, 1621.5 and 1070.8 cm-1, respectively. On the other hand, we observed mJ |±9/2〉 as the ground state for [Dy(C7H7)2]3- (4) and [Dy(C8H8)2]- (5) complexes with significantly smaller Ucal values of 237.1 and 38.6 cm-1 respectively. For the nine-membered ring [Dy(C9H9)2]+ (6) complex, we observed the stabilization of the mJ |±1/2〉 ground state, with the first excited state being located ∼29 cm-1 higher in energy. AILFT-NEVPT2 ligand field splitting analysis indicates that the presence of π-type 4f-ligand interactions in complexes 1-3 help generate the axial-ligand field, while the δ-type interactions in complexes 4-5 generate the equatorial ligand field despite the ligands approaching from the axial direction. As the ring size increases, φ-type interactions dominate, generating a pure equatorial ligand field stabilising mJ |±1/2〉 as the ground state for 6. Calculations suggest that the nature of the ligand field mainly governs the Ucal values in the following order: 4f-Lσ > 4f-Lπ > 4f-Lδ > 4f-Lφ. Calculations were performed by replacing ligands with CHELPG charges to access the crystal field (CF) effects which suggests the stabilization of pure mJ |±15/2〉 in all the charge-embedded models (1Q-6Q). Our findings point out that the crystal field and ligand field effects complement each other and generate a giant barrier for magnetic relaxation in the small ring complexes 1-3, while a relatively weak crystal field and adverse 4f-Lδ/4f-Lφ interactions diminish the SMM behaviour in the large ring complexes 4-6.

Purification of the Transmembrane Polypeptide Channel Complex of the Salmonella Flagellar Type III Secretion System.

Many motile bacteria employ the flagellar type III secretion system (fT3SS) to build the flagellum on the cell surface. The fT3SS consists of a transmembrane export gate complex, which acts as a proton/protein antiporter that couples proton flow with flagellar protein export, and a cytoplasmic ATPase ring complex, which works as an activator of the export gate complex. Three transmembrane proteins, FliP, FliQ, and FliR, form a core structure of the export gate complex, and this core complex serves as a polypeptide channel that allows flagellar structural subunits to be translocated across the cytoplasmic membrane. Here, we describe the methods for overproduction, solubilization, and purification of the Salmonella FliP/FliQ/FliR complex.

The ring saga: looking back at the discovery of γ-tubulin and γ-tubulin ring complexes

For many years, two central, unanswered questions in cytoskeleton research were how microtubule assembly is nucleated and microtubule polarity established. The discoveries of γ-tubulin and γ-tubulin ring complexes were key advances that allowed these questions to be substantially answered. The discovery of γ-tubulin was the product of a genetic screen in Aspergillus nidulans for genes important for microtubule function. γ-Tubulin is a member of the tubulin superfamily of proteins, closely related to α- and β-tubulin but distinct from both. It is ubiquitous in eukaryotes, and in many organisms there are small families of γ-tubulin genes. γ-Tubulin and associated proteins form ring-like complexes that localize to microtubule-organizing centers (MTOCs) and play an important role in the nucleation of microtubule assembly from MTOCs and the establishment of microtubule polarity.

Magma fractionation and emplacement mechanism in a subvolcanic plumbing system in a continental region: constraints from the late Neoproterozoic Wadi Dib ring complex in the Eastern Desert, Egypt

Magma fractionation and emplacement mechanism in a subvolcanic plumbing system in a continental region: constraints from the late Neoproterozoic Wadi Dib ring complex in the Eastern Desert, Egypt

Collective action and shellfish harvesting practices among Late Archaic villagers of the South Atlantic Bight

Indigenous coastal communities across the globe sustainably harvested oysters and other shellfish species for millennia. European colonialism and the emergence of market-based institutions, however, lead to the eventual demise of many oyster reefs and fisheries beginning in the late 1800 s. Circular shell rings situated on Georgia’s South Atlantic coast are the preserved remnants of Native American village communities during the Late Archaic (5000–3000 cal. BP). Mollusk shells from these archaeological contexts hold chemical clues into past human-environmental interactions and thus give insight into Indigenous histories and sustainable shellfish harvesting practices. In this paper, we interpret shellfish geochemistry data (oxygen isotopes, δ18O) from the Sapelo Island Shell Ring Complex within a theoretical framework of cooperation and collective action to understand the ways in which Ancestral Muskogean people of Sapelo Island, Georgia, effectively managed and sustained oyster reefs and other coastal fisheries during the Late Archaic. More specifically, δ18O values from 18 oysters and 57 clams were used to determine season of harvest and to estimate salinity values of the habitats from which the shells were harvested. Results demonstrate considerable variation in estimated salinity values and some statistically significant differences in δ18O and salinity values between shells harvested in different seasons. This indicates that the sedentary villagers who lived at the Sapelo Shell Ring Complex were moving around seasonally and using an array of habitats. We argue that this suggests the presence of social institutions or rules that governed the use of coastal estuaries so that mollusks were not overexploited.

The conserved centrosomin motif, γTuNA, forms a dimer that directly activates microtubule nucleation by the γ-tubulin ring complex (γTuRC).

To establish the microtubule cytoskeleton, the cell must tightly regulate when and where microtubules are nucleated. This regulation involves controlling the initial nucleation template, the γ-tubulin ring complex (γTuRC). Although γTuRC is present throughout the cytoplasm, its activity is restricted to specific sites including the centrosome and Golgi. The well-conserved γ-tubulin nucleation activator (γTuNA) domain has been reported to increase the number of microtubules (MTs) generated by γTuRCs. However, previously we and others observed that γTuNA had a minimal effect on the activity of antibody-purified Xenopus γTuRCs in vitro (Thawani et al., eLife, 2020; Liu et al., 2020). Here, we instead report, based on improved versions of γTuRC, γTuNA, and our TIRF assay, the first real-time observation that γTuNA directly increases γTuRC activity in vitro, which is thus a bona fide γTuRC activator. We further validate this effect in Xenopus egg extract. Via mutation analysis, we find that γTuNA is an obligate dimer. Moreover, efficient dimerization as well as γTuNA's L70, F75, and L77 residues are required for binding to and activation of γTuRC. Finally, we find that γTuNA's activating effect opposes inhibitory regulation by stathmin. In sum, our improved assays prove that direct γTuNA binding strongly activates γTuRCs, explaining previously observed effects of γTuNA expression in cells and illuminating how γTuRC-mediated microtubule nucleation is regulated.

Detection and Analysis of Microtubule Nucleator γ-Tubulin Ring Complex.

Golgi-derived microtubules constitute an asymmetrical microtubule network that drives polarized transport of vesicles to support cell polarization and directional migration. Golgi-based microtubule nucleation requires the γ-tubulin ring complex (γTuRC), the principal microtubule nucleator in animal cells. In this chapter, we present methods for detecting γTuRC components and associated proteins on the Golgi, examining Golgi-based microtubule nucleation, and measuring the microtubule-nucleating activity of isolated γTuRCs. These approaches have been demonstrated to be effective for assessing the microtubule-organizing function of the Golgi complex.