Triad Formation Research Articles

A Holistic Analysis of Alzheimer’s Disease-Associated lncRNA Communities Reveals Enhanced lncRNA-miRNA-RBP Regulatory Triad Formation Within Functionally Segregated Clusters

Recent studies on the regulatory networks implicated in Alzheimer’s disease (AD) evince long non-coding RNAs (lncRNAs) as crucial regulatory players, albeit a poor understanding of the mechanism. Analyzing differential gene expression in the RNA-seq data from the post-mortem AD brain hippocampus, we categorized a list of AD-dysregulated lncRNA transcripts into functionally similar communities based on their k-mer profiles. Using machine-learning-based algorithms, their subcellular localizations were mapped. We further explored the functional relevance of each community through AD-dysregulated miRNA, RNA-binding protein (RBP) interactors, and pathway enrichment analyses. Further investigation of the miRNA–lncRNA and RBP–lncRNA networks from each community revealed the top RBPs, miRNAs, and lncRNAs for each cluster. The experimental validation community yielded ELAVL4 and miR-16-5p as the predominant RBP and miRNA, respectively. Five lncRNAs emerged as the top-ranking candidates from the RBP/miRNA-lncRNA networks. Further analyses of these networks revealed the presence of multiple regulatory triads where the RBP–lncRNA interactions could be augmented by the enhanced miRNA–lncRNA interactions. Our results advance the understanding of the mechanism of lncRNA-mediated AD regulation through their interacting partners and demonstrate how these functionally segregated but overlapping regulatory networks can modulate the disease holistically.Graphical

Stabilization of the Protein Structure by the Many-Body Cooperative Effect in the NH/π Hydrogen-bonding Tryptophan Triad.

The indole ring of tryptophan can form NH/π hydrogen bonds, acting both as a hydrogen donor at the NH group in the pyrrole subring and as a hydrogen acceptor at the benzene subring. In the structural core of the trimeric stable protein Pholiota squarrosa lectin (PhoSL), three indoles are symmetrically arranged and form NH/π hydrogen bonds among each other. Here, we conducted quantum chemical calculations on this indole triad by using various methods and basis sets. The analyses revealed cooperativity in triad formation, with the many-body effect contributing approximately -2 kcal mol-1, which significantly stabilizes this protein. Symmetry-adapted perturbation theory ascribed this effect to the induced polarization. The electrostatic potential and atomic charges indeed revealed a charge redistribution through the NH/π hydrogen bond, which was favorable for triad formation.

Intratumoral immune triads are required for immunotherapy-mediated elimination of solid tumors

Tumor-specific CD8+ Tcells are frequently dysfunctional and unable to halt tumor growth. We investigated whether tumor-specific CD4+ Tcells can be enlisted to overcome CD8+ Tcell dysfunction within tumors. We find that the spatial positioning and interactions of CD8+ and CD4+ Tcells, but not their numbers, dictate anti-tumor responses in the context of adoptive Tcell therapy as well as immune checkpoint blockade (ICB): CD4+ Tcells must engage with CD8+ Tcells on the same dendritic cell during the effector phase, forming a three-cell-type cluster (triad) to license CD8+ Tcell cytotoxicity and cancer cell elimination. When intratumoral triad formation is disrupted, tumors progress despite equal numbers of tumor-specific CD8+ and CD4+ Tcells. Inpatients with pleural mesothelioma treated with ICB, triads are associated with clinical responses. Thus,CD4+ Tcells and triads are required for CD8+ Tcell cytotoxicity during the effector phase and tumor elimination.

Integrated multi-omics approach reveals the role of striated muscle preferentially expressed protein kinase in skeletal muscle including its relationship with myospryn complex.

Autosomal-recessive mutations in SPEG (striated muscle preferentially expressed protein kinase) have been linked to centronuclear myopathy with or without dilated cardiomyopathy (CNM5). Loss of SPEG is associated with defective triad formation, abnormal excitation-contraction coupling, calcium mishandling and disruption of the focal adhesion complex in skeletal muscles. To elucidate the underlying molecular pathways, we have utilized multi-omics tools and analysis to obtain a comprehensive view of the complex biological processes and molecular functions. Skeletal muscles from 2-month-old SPEG-deficient (Speg-CKO) and wild-type (WT) mice were used for RNA sequencing (n=4 per genotype) to profile transcriptomics and mass spectrometry (n=4 for WT; n=3 for Speg-CKO mice) to profile proteomics and phosphoproteomics. In addition, interactomics was performed using the SPEG antibody on pooled muscle lysates (quadriceps, gastrocnemius and triceps) from WT and Speg-CKO mice. Based on the multi-omics results, we performed quantitative real-time PCR, co-immunoprecipitation and immunoblot to verify the findings. We identified that SPEG interacts with myospryn complex proteins CMYA5, FSD2 and RyR1, which are critical for triad formation, and that SPEG deficiency results in myospryn complex abnormalities (protein levels decreased to 22±3% for CMYA5 [P<0.05] and 18±3% for FSD2 [P<0.01]). Furthermore, SPEG phosphorylates RyR1 at S2902 (phosphorylation level decreased to 55±15% at S2902 in Speg-CKO mice; P<0.05), and its loss affects JPH2 phosphorylation at multiple sites (increased phosphorylation at T161 [1.90±0.24-fold], S162 [1.61±0.37-fold] and S165 [1.66±0.13-fold]; decreased phosphorylation at S228 and S231 [39±6%], S234 [50±12%], S593 [48±3%] and S613 [66±10%]; P<0.05 for S162 and P<0.01 for other sites). On analysing the transcriptome, the most dysregulated pathways affected by SPEG deficiency included extracellular matrix-receptor interaction (P<1e-15) and peroxisome proliferator-activated receptor signalling (P<9e-14). We have elucidated the critical role of SPEG in the triad as it works closely with myospryn complex proteins (CMYA5, FSD2 and RyR1), it regulates phosphorylation levels of various residues in JPH2 and S2902 in RyR1, and its deficiency is associated with dysregulation of several pathways. The study identifies unique SPEG-interacting proteins and their phosphorylation functions and emphasizes the importance of using a multi-omics approach to comprehensively evaluate the molecular function of proteins involved in various genetic disorders.

Network Evolution Model with Preferential Attachment at Triadic Formation Step

It is recognized that most real systems and networks exhibit a much higher clustering with comparison to a random null model, which can be explained by a higher probability of the triad formation—a pair of nodes with a mutual neighbor have a greater possibility of having a link between them. To catch the more substantial clustering of real-world networks, the model based on the triadic closure mechanism was introduced by P. Holme and B. J. Kim in 2002. It includes a “triad formation step” in which a newly added node links both to a preferentially chosen node and to its randomly chosen neighbor, therefore forming a triad. In this study, we propose a new model of network evolution in which the triad formation mechanism is essentially changed in comparison to the model of P. Holme and B. J. Kim. In our proposed model, the second node is also chosen preferentially, i.e., the probability of its selection is proportional to its degree with respect to the sum of the degrees of the neighbors of the first selected node. The main goal of this paper is to study the properties of networks generated by this model. Using both analytical and empirical methods, we show that the networks are scale-free with power-law degree distributions, but their exponent γ is tunable which is distinguishable from the networks generated by the model of P. Holme and B. J. Kim. Moreover, we show that the degree dynamics of individual nodes are described by a power law.

Triad resonance of flexural gravity waves in the presence of shear current with constant vorticity

This study examines the formation of triads of flexural gravity wave in a homogeneous fluid within the context of blocking dynamics due to the presence of shear current. This study will enable us to understand the distribution of wave energy on an ice-covered sea surface. New classes of triads for flexural gravity waves are introduced depending on the direction of wave propagation with following and opposing currents. The study reveals that triad formation occurs due to the interaction of flexural gravity waves irrespective of the presence of compression and current, which has not been found in the case of free surface gravity waves. In addition, at most, three triads are formed in the case of flexural gravity waves in the presence of following and opposing currents prior to the threshold of blocking. In contrast, at least three triads are formed for any frequency within the primary and secondary blocking limits for certain values of compressive force and current speed. On the other hand, 11 triads are formed in the presence of uniform current speed as well as in the case of linear shear current with constant vorticity for a certain frequency within the blocking limit for higher values of compressive force and current speed.

Caveolae and Bin1 form ring-shaped platforms for T-tubule initiation.

Excitation-contraction coupling requires a highly specialized membrane structure, the triad, composed of a plasma membrane invagination, the T-tubule, surrounded by two sarcoplasmic reticulum terminal cisternae. Although the precise mechanisms governing T-tubule biogenesis and triad formation remain largely unknown, studies have shown that caveolae participate in T-tubule formation and mutations of several of their constituents induce muscle weakness and myopathies. Here, we demonstrate that, at the plasma membrane, Bin1 and caveolae composed of caveolin-3 assemble into ring-like structures from which emerge tubes enriched in the dihydropyridine receptor. Bin1 expression lead to the formation of both rings and tubes and we show that Bin1 forms scaffolds on which caveolae accumulate to form the initial T-tubule. Cav3 deficiency caused by either gene silencing or pathogenic mutations results in defective ring formation and perturbed Bin1-mediated tubulation that may explain defective T-tubule organization in mature muscles. Our results uncover new pathophysiological mechanisms that may prove relevant to myopathies caused by Cav3 or Bin1 dysfunction.

Endosperm Development Traits in a Comparative Analysis of Endospermogenesis and Embryogenesis in Angiosperms.

The article discusses the fertilization process, the nucleus position in the primary cell, specifics of early endosperm development, and the principles of its classification. A new, refined classification was proposed for endosperm development modes to include three hierarchic levels: types, subtypes, and variations. Two types were distinguished by the morphogenetic potentials of the micropylar and chalazal primary cells: cellular (karyokinesis is completed with cytokinesis in both cells) and helobial (only karyokinesis takes place in both cells, and the chalazal cell sometimes remains mononucleate). The nuclear endosperm was considered as a subtype of the helobial type. Subtypes were isolated by the extent to which the micropylar and chalazal cells are involved in forming the endosperm. Variations were recognized within the subtypes by the position of walls during the tetrad or triad formation in the cellular endosperm or the number of nuclei in the chalazal cell in the helobial endosperm. The types of embryogenesis are possible to compare with subtypes or even variations of the cellular endosperm type in a comparative analysis of flowering plants, and both of the traits (the contribution of micropylar and chalazal cell derivatives to the endosperm formation and the pattern of primary cell division with the form of the tetrad) should be considered together. Two subtypes and two variations are possible to consider for the helobial endosperm.

Self-assembling cobalt(II) porphyrin - fullero [60]pyrrolidine triads. Synthesis and spectral properties in the ground and excited state

Meso‑4-Trifluoromethylphenyl substituted cobalt(II)porphin, CoT(CF3P)P, and its coordination 1: 2 complex (triad) with 1-N-methyl-2-(pyridin-4-yl)-3,4-fullero [60]pyrrolidine, (PyC60)2CoT(CF3P)P, were firstly synthesized and the properties of their molecular solutions were studied. The chemical structure of CoT(CF3P)P and triad was fully confirmed by the UV–vis, IR and 1H NMR spectroscopy and mass spectrometry methods. The triad formation mechanism as the two-step equilibrium characterized by equilibrium constant K1 (5.89 ± 1.50) × 104 M−1 and K2 (7.40 ± 1.66) × 105 M−1 was revealed. Using femtosecond pump-probe spectroscopy measurements, the photoinduced electron transfer in the synthesized triad was described. The excited state lifetime of the precursors, CoTPP, CoT(CF3P)P and the charge separated state lifetime of (PyC60)2CoT(CF3P)P compared with unsubstituted (PyC60)2CoTPP were determined. It was established that the CF3 groups in the porphyrin periphery slightly change the CoTPP/CoT(CF3P)P excited state lifetime and the electron transfer rate constant in the correspondent triads.

Antioxidant thioether core-crosslinked nanoparticles prevent the bilateral spread of secondary injury to protect spatial learning and memory in a controlled cortical impact mouse model of traumatic brain injury

The secondary phase of traumatic brain injury (TBI) is partly caused by the release of excess reactive oxygen species (ROS) from the primary injury. However, there are currently no therapies that have been shown to reduce the secondary spread of injury beyond the primary insult. Nanoparticles offer the ability to rapidly accumulate and be retained in injured brain for improved target engagement. Here, we utilized systemically administered antioxidant thioether core-cross-linked nanoparticles (NP1) that scavenge and inactivate ROS to reduce this secondary spread of injury in a mild controlled cortical impact (CCI) mouse model of TBI. We found that NP1 treatment protected CCI mice from injury induced learning and memory deficits observed in the Morris water maze (MWM) test at 1-month post-CCI. This protection was likely a result of NP1-mediated reduction in oxidative stress in the ipsilateral hemisphere as determined by immunofluorescence imaging of markers of oxidative stress and the spread of neuroinflammation into the contralateral hippocampus as determined by immunofluorescence imaging of activated microglia and neuron-astrocyte-microglia triad formation. These data suggest NP1-mediated reduction in post-traumatic oxidative stress correlates with the reduction in the spread of injury to the contralateral hippocampus to protect spatial memory and learning in CCI mice. Therefore, these materials may offer an improved treatment strategy to reduce the secondary spread of TBI.

Self-organizing donor-acceptor assemblies of cobalt(II) porphyrin ligated with gold(III) porphyrin or fullero[60]pyrrolidine in liquid medium

This work provides the results of the spectroscopic, electrochemical, and photoelectrochemical study of the self-assembly in the systems consisting of (2,3,7,8,12,13,17,18-octaethylporphinato)cobalt(II) (CoOEP), (2,3,7,8,12,18-hexamethyl,13,17-diethyl,5-(4-pyridyl)porphinato)gold(III) chloride (ClAuP)/1′-N-methyl-2′-(pyridin-4-yl)pyrrolidino[3′,4′:1,2][60]fullerene (PyC60), and 1,2-dichlorobenzene/toluene as an electron donor, an acceptor, and a liquid medium, respectively. The results indicate axial coordination of electron acceptor functionalized by the pyridine units by cobalt(II) porphyrin with the donor-acceptor (ClAuP)2CoOEP/(PyC60)2CoOEP triad formation. The thermodynamic stability constant is (4.91 ± 1.10)×109 L2 mol−2 and (4.10 ± 1.14) ×1010 L2 mol−2 for ClAuP and PyC60, respectively. The formation of supramolecules was confirmed by UV, visible, IR, 1H NMR spectroscopy and cyclic voltammetry. DFT calculations (B3LYP+def2-SVP) detect octahedral geometry of the coordination center with the trans configuration of the axial ligands in the triads as well as the excited state calculations establish photoinduced electron transfer (PET). The jphavg value 266 μA cm−2 for the Ti|(ClAuP)2CoOEP|0.5 M Na2SO4|Pt system being 2–2.5 times higher compared with the precursors and slightly higher compared with (PyC60)2CoOEP shows that gold(III) porphyrin as an electron acceptor can be successfully used. The development of donor/acceptor platform selection strategy for the obtaining of photoactive supramolecules is an urgent task for future deriving of photovoltaic devices.

Striated Preferentially Expressed Protein Kinase (SPEG)-Deficient Skeletal Muscles Display Fewer Satellite Cells with Reduced Proliferation and Delayed Differentiation

Centronuclear myopathies (CNMs) are a subtype of congenital myopathies characterized by skeletal muscle weakness and an increase in the number of central myonuclei. SPEG (striated preferentially expressed protein kinase) has been identified as the sixth gene associated with CNM, and it has been shown that striated muscle-specific Speg-knockout (KO) mice have defective triad formation, abnormal excitation-contraction coupling, and calcium mishandling. The impact of SPEG deficiency on the survival and function of myogenic cells remains to be deciphered. In this study, the authors examined the overall population, proliferation, and differentiation of myogenic cells obtained from striated muscle-specific Speg-KO mice and compared them with wild-type (WT) controls. SPEG-deficient skeletal muscles contained fewer myogenic cells, which on further study demonstrated reduced proliferation and delayed differentiation compared with those from WT muscles. Regenerative response to skeletal muscle injury in Speg-KO mice was compared with that of WT mice, leading to the identification of similar abnormalities including fewer satellite cells, fewer dividing cells, and an increase in apoptotic cells in KO mice. Overall, these results reveal specific abnormalities in myogenic cell number and behavior associated with SPEG deficiency. Similar satellite cell defects have been reported in mouse models of MTM1- and DNM2-associated CNM, suggestive of shared underlying pathways.

De novo phosphoinositide synthesis in zebrafish is required for triad formation but not essential for myogenesis.

Phosphoinositides (PIPs) and their regulatory enzymes are key players in many cellular processes and are required for aspects of vertebrate development. Dysregulated PIP metabolism has been implicated in several human diseases, including a subset of skeletal myopathies that feature structural defects in the triad. The role of PIPs in skeletal muscle formation, and particularly triad biogenesis, has yet to be determined. CDP-diacylglycerol-inositol 3-phosphatidyltransferase (CDIPT) catalyzes the formation of phosphatidylinositol, which is the base of all PIP species. Loss of CDIPT should, in theory, result in the failure to produce PIPs, and thus provide a strategy for establishing the requirement for PIPs during embryogenesis. In this study, we generated cdipt mutant zebrafish and determined the impact on skeletal myogenesis. Analysis of cdipt mutant muscle revealed no apparent global effect on early muscle development. However, small but significant defects were observed in triad size, with T-tubule area, inter terminal cisternae distance and gap width being smaller in cdipt mutants. This was associated with a decrease in motor performance. Overall, these data suggest that myogenesis in zebrafish does not require de novo PIP synthesis but does implicate a role for CDIPT in triad formation.

Combined effects of amplitude, frequency and bandwidth on wavepackets in laminar turbulent transition

This study concerns wavepackets in laminar turbulent transition in a Blasius boundary layer. While initial amplitude and frequency have well-recognized roles in the transition process, the current study on the combined effects of amplitude, frequency, and bandwidth on the propagation of wavepackets is believed to be new. Because of the complexity of the system, these joint variations in multiple parameters could give rise to effects not seen through the variation of any single parameter. Direct numerical simulations (DNS) are utilized in a full factorial (fully crossed) design to investigate both individual and joint effects of variation in the simulation parameters, with a special focus on three distinct variants of wavepacket transition—the reverse Craik triad formation sequence, concurrent N-type and K-type transition and abrupt shifts in dominant frequency. From our factorial study, we can summarize the key transition trends of wavepackets as follows:1.Broad bandwidth wavepackets predominantly transit to turbulence via the N-route. This tendency remains strong even as the frequency width is reduced.2.Narrow bandwidth wavetrains exhibit predominantly K-type transition. The front broadband part of an emerging wavetrain may experience N-type transition, but this wavefront should not be considered as a part of truly narrow-bandwidth wavepackets.3.K-type transition is the most likely for wavepackets that are initiated with high energy/amplitude and/or with the peak frequency at the lower branch of the neutral stability curve.

Structure properties of collaboration network with tunable clustering

A collaboration network model with improved triad formation (ITF) step is proposed and denominated as one-mode collaboration network with tunable clustering. We systematically study its structural features and find this network model can much more accurately reflect cooperative relationship in real society. We acquire expression of nodes degree distribution P(k) that possesses a power-law form with exponent γ whose range is (1,3]. Expression of clustering coefficient C(k) for nodes with degree k is also obtained and composes of a power-law section and a constant section. We verify clustering coefficient of individual nodes increases monotonically with the increase of parameter p that is the probability of ITF step occurring. When specific forms of P(k) and C(k) are given, the model’s average clustering coefficient C can be derived approximately. Finally, we analytically testify average path length L(N) increases with network size N in accordance with logarithmic form. When N is fixed, average path length of entire network decreases with the increase of p. All theoretically predicted results are consistent with simulative experimental results.

The hepatic architecture of the coelacanth differs from that of the lungfish in portal triad formation.

The liver architecture of vertebrates can be classified into two types, the portal triad type (having periportal bile ducts) and the non-portal triad type (having non-periportal bile ducts). The former is detectable in the tetrapod liver whereas the lungfish liver has the latter. It remains to be revealed which type of hepatic architecture the coelacanth, which together with the lungfish belongs to the Sarcopterygii, possesses. The present study was undertaken to determine the histological characteristics of the coelacanth liver, and to compare with those of other vertebrates. The coelacanth liver had periportal bile ducts and ductules as detected in mammalian livers. The hepatic artery was found around large portal veins. Hagfish, shark, bichir, sturgeon, bowfin and frog livers had periportal bile ducts and bile ductules, whereas most intrahepatic bile ducts of the lungfish were independent of the distribution of the portal veins as seen in the Otocephala and Euteleostei. The lungfish liver developed duct and ductule structures in the parenchyma. These data indicate that the coelacanth liver had a mammalian-type hepatic architecture with a portal triad, and that the ancestors of tetrapods may have had a portal triad-type liver architecture.

Exploiting reciprocity toward link prediction

This paper addresses link prediction problem in directed networks by exploiting reciprocative nature of human relationships. It first proposes a null model to present evidence that reciprocal links influence the process of “triad formation”. Motivated by this, reciprocal links are exploited to enhance link prediction performance in three ways: (a) a reciprocity-aware link weighting technique is proposed, and existing weighted link prediction methods are applied over the resultant weighted network; (b) new link prediction methods are proposed, which exploit reciprocity; and (c) existing and proposed methods are combined toward supervised prediction to enhance the prediction performance further. All experiments are carried out on two real directed network datasets.

Cobalt(II) porphyrin axially coordinated with 2′-(pyridin-4-yl)-5′-(pyridin-2-yl)-1′-(pyridin-2-ylmethyl)-2′,4′-dihydro-1′H-pyrrolo[3′,4′ : 1,2](C60-Ih)[5,6]fullerene: formation, chemical structure, and spectroscopic properties

Construction of new effective photovoltaic devices based on organic dyes has important implications for modern and future technologies. In this article, we studied the equilibrium, the rate, and the spectral manifestation of the reaction of [(2,3,7,8,12,18-hexamethyl,13,17-diethyl,5-(2-pyridyl)porphyrinato)cobalt(II)]–[2′-(pyridin-4-yl)-5′-(pyridin-2-yl)-1′-(pyridin-2-ylmethyl)-2′,4′-dihydro-1′H-pyrrolo[3′,4′ : 1,2](C60-Ih)[5,6]fullerene] triad formation as well as its spectral properties and photo electrochemical behavior. The cobalt porphyrin–pyridyl-substituted fullerene mixtures in toluene are self-assembling systems due to axial donor–acceptor binding between Co of the porphyrin complex and N-pyridyl of the substituted fullerene. The formation rate constant, k298K, and the stability constant, K298K, of donor–acceptor triad formed by coordination of two substituted fullerene molecules to Co porphyrin are (44.4 ± 0.8) mol L−1 s−1 and (56 ± 16)×107 L2 mol−2, respectively. Modification of the titanium electrode coated with the natural oxide film was carried out using the porphyrin–fullerene triad and its individual components. Photopotential and photocurrent density of the system with modified electrode were studied. The obtained results are of interest for creating porphyrin-based donor–acceptor systems as components in organic photovoltaics.

Topology evolution model for ad hoc‐cellular hybrid networks based on complex network theory

SummarySwitching and routing are two important issues in the ad hoc‐cellular hybrid network. Frequently switching between different network modes will lead to the increase of time delay, and unreasonable routing will reduce information transmission efficiency and shorten the network life. In order to reduce the influence of these problems to the network, in this paper, we proposed a robust and efficient ad hoc network topology evolution model, which can reduce the switching probability and establish efficient transmission paths. In this model, the mobile users' residual energy, available channel quality, and the importance are taken into account. The model is based on Barrat, Barthelemy, and Vespignani model and the triad formation mechanism, which ensures the network topology have scale‐free and small‐world features. The topology construction process of the ad hoc network is composed of three parts: (i) new users join the network; (ii) new users establish connections with existing users; (iii) the connections between existing users were deleted because of the network optimization and the interference of cellular network. Simulation results show that the network built by this model with small‐world and scale‐free features has small average shortest path length and the robustness against random nodes failure, which will significantly improve the transmission efficiency and reduce the probability of switching between different transmission model. Copyright © 2016 John Wiley &amp; Sons, Ltd.

The Microtubule-Associated Protein CLIMP-63 is a New Member of the Calcium Release Complex

Muscle contraction is achieved when an efficient excitation signal at the plasma membrane triggers intracellular calcium release. This process called “excitation-contraction (E-C) coupling” relies on a macromolecular protein complex, spanning the plasma membrane and the sarcoplasmic reticulum (SR), containing the calcium channel of the SR, the ryanodine receptor (RyR). This calcium release complex is present exclusively in highly organized membrane structures called triads. A triad is composed of two SR terminal cisternae surrounding a plasma membrane transverse-tubule. This architecture is essential to sustain the activity of the calcium channel RyR1, which is located in the membrane of SR terminal cisternae. However, little is known about the molecular mechanisms allowing the formation and maintenance of SR terminal cisternae. Triadin is a member of this complex, present in the SR membrane and interacting with RyR1. Deletion of the triadin gene leads to partial disorganisation of SR membranes in skeletal muscles, with abnormal orientation of part of the triads. Triadin could play a role in the structure of sarcoplasmic reticulum to allow efficient E-C coupling. We have shown that triadin could indirectly interact with the microtubules, and therefore anchor the sarcoplasmic reticulum to the microtubule network (Fourest-Lieuvin, J.Cell.Science, 2012). Using mass spectrometry analysis of the proteins co-immunoprecipitated with triadin, we have identified a new partner of triadin, CLIMP-63 which could be involved in this function. CLIMP-63 is a shaping protein able to mediate the anchoring of the reticulum to microtubules. CLIMP-63 has been involved in the shaping of endoplasmic reticulum in neurons (Cui-Wang, Cell, 2012). We have dissected the interacting domains between CLIMP-63 and triadin, and studied the consequences of this association for muscle function, and triad formation or maintenance.