Expression of intermediate filament proteins during development of Xenopus laevis. II. Identification and molecular characterization of desmin

Development ◽  
1989 ◽  
Vol 105 (2) ◽  
pp. 299-307 ◽  
Author(s):  
H. Herrmann ◽  
B. Fouquet ◽  
W.W. Franke

During embryogenesis of avian and mammalian species the formation of intermediate filaments (IFs) containing desmin is characteristic for myogenesis. In view of important differences of patterns of IF protein expression in embryogenic pathways of amphibia on the one hand and birds and mammals on the other, we have decided to study the expression of desmin during early embryogenesis of Xenopus laevis by cDNA hybridization and antibody reactions. Here we describe the isolation of a cDNA clone encoding Xenopus desmin and the deduced amino acid sequence (458 residues; Mr 52,800) which displays a very high degree of conservation during vertebrate evolution from Xenopus to chicken and hamster, with a similar degree of sequence divergence between all three species compared. In addition, we have noted, by both cDNA-hybrid-selection-translation and immunoblotting of cytoskeletal proteins a second desmin-related polypeptide of Mr approximately 49,000. RNA (Northern) blot analyses show the occurrence of three different desmin mRNAs (1.9, 2.6 and 3.0 kb) which seem to represent different polyadenylation sites, displaying quantitative differences in different kinds of muscle tissues. During embryogenesis, desmin mRNA has first been detected in stage-14 embryos and then increases drastically to high levels at stage 18 and thereafter. Immunofluorescence microscopy using desmin-specific antibodies shows that this synthesis of desmin is restricted to somite tissue. The embryonic time course of synthesis of desmin and desmin mRNA is discussed in relation to those of other muscle proteins.

Development ◽  
1989 ◽  
Vol 105 (2) ◽  
pp. 279-298
Author(s):  
H. Herrmann ◽  
B. Fouquet ◽  
W.W. Franke

To provide a basis for studies of the expression of genes encoding the diverse kinds of intermediate-filament (IF) proteins during embryogenesis of Xenopus laevis we have isolated and characterized IF protein cDNA clones. Here we report the identification of two types of Xenopus vimentin, Vim1 and Vim4, with their complete amino acid sequences as deduced from the cloned cDNAs, both of which are expressed during early embryogenesis. In addition, we have obtained two further vimentin cDNAs (Vim2 and 3) which are sequence variants of closely related Vim1. The high evolutionary conservation of the amino acid sequences (Vim1: 458 residues; Mr approximately 52,800; Vim4: 463 residues; Mr approximately 53,500) to avian and mammalian vimentin and, to a lesser degree, to desmin from the same and higher vertebrate species, is emphasized, including conserved oligopeptide motifs in their head domains. Using these cDNAs in RNA blot and ribonuclease protection assays of various embryonic stages, we observed a dramatic increase of vimentin RNA at stage 14, in agreement with immunocytochemical results obtained with antibody VIM-3B4. The significance of very weak mRNA signals detected in earlier stages is discussed in relation to negative immunocytochemical results obtained in these stages. The first appearance of vimentin has been localized to a distinct mesenchymal cell layer underlying the neural plate or tube, respectively. The results are discussed in relation to programs of de novo synthesis of other cytoskeletal proteins in amphibian and mammalian development.


2021 ◽  
Author(s):  
Evan Couser ◽  
Jennifer L Chlebek ◽  
Ankur B Dalia

Bacterial surface appendages called type IVa pili (T4aP) promote diverse activities including DNA uptake, twitching motility, and virulence. These activities rely on the ability of T4aP to dynamically extend and retract from the cell surface. Dynamic extension relies on a motor ATPase commonly called PilB. Most T4aP also rely on specific motor ATPases, commonly called PilT and PilU, to dynamically and forcefully retract. Here, we systematically assess whether motor ATPases from 4 distinct T4aP could functionally complement Vibrio cholerae mutants that lacked their endogenous motors. We found that the retraction ATPases PilT and PilU are highly promiscuous and promote retraction of the V. cholerae competence T4aP despite a high degree of sequence divergence. In contrast, orthologous extension ATPases were not able to mediate extension of the V. cholerae competence T4aP despite a similar degree of sequence divergence. Also, we show that one of the PilT orthologs characterized does not support PilU-dependent retraction and we attributed this loss of activity to the 3' end of the gene, which suggests that the C-terminus of PilT plays an important role in promoting PilU-dependent retraction. Together, our data suggest that retraction ATPases have maintained a high degree of promiscuity for promoting retraction of diverse T4aP, while extension ATPases have evolved to become highly specific for their cognate systems.


1989 ◽  
Vol 109 (4) ◽  
pp. 1695-1709 ◽  
Author(s):  
K Schlaeppi ◽  
J Deflorin ◽  
T Seebeck

The flagellum of the parasitic hemoflagellate Trypanosoma brucei contains two major structures: (a) the microtubule axoneme, and (b) a highly ordered, filamentous array, the paraflagellar rod (PFR). This is a complex, three-dimensional structure, of yet unknown function, that extends along most of the axoneme and is closely linked to it. Its major structural component is a single protein of 600 amino acids. This PFR protein can assume two different conformations, resulting in two distinct bands of apparent molecular masses of 73 and 69 kD in SDS-gel electrophoresis. Secondary structure predictions indicate a very high helix content. Despite its biochemical similarity to the intermediate filament proteins (solubility properties, amino acid composition, and high degree of helicity), the PFR protein does not belong in this class of cytoskeletal proteins. The PFR protein is coded for by two tandemly linked genes of identical nucleotide sequence. Both genes are transcribed into stable mRNAs of very similar length that carry the mini-exon sequence at their 5' termini.


2016 ◽  
Vol 114 (2) ◽  
pp. E191-E199 ◽  
Author(s):  
Elisa D’Este ◽  
Dirk Kamin ◽  
Francisco Balzarotti ◽  
Stefan W. Hell

We used stimulated emission depletion (STED) superresolution microscopy to analyze the nanoscale organization of 12 glial and axonal proteins at the nodes of Ranvier of teased sciatic nerve fibers. Cytoskeletal proteins of the axon (betaIV spectrin, ankyrin G) exhibit a high degree of one-dimensional longitudinal order at nodal gaps. In contrast, axonal and glial nodal adhesion molecules [neurofascin-186, neuron glial-related cell adhesion molecule (NrCAM)] can arrange in a more complex, 2D hexagonal-like lattice but still feature a ∼190-nm periodicity. Such a lattice-like organization is also found for glial actin. Sodium and potassium channels exhibit a one-dimensional periodicity, with the Nav channels appearing to have a lower degree of organization. At paranodes, both axonal proteins (betaII spectrin, Caspr) and glial proteins (neurofascin-155, ankyrin B) form periodic quasi–one-dimensional arrangements, with a high degree of interdependence between the position of the axonal and the glial proteins. The results indicate the presence of mechanisms that finely align the cytoskeleton of the axon with the one of the Schwann cells, both at paranodal junctions (with myelin loops) and at nodal gaps (with microvilli). Taken together, our observations reveal the importance of the lateral organization of proteins at the nodes of Ranvier and pave the way for deeper investigations of the molecular ultrastructural mechanisms involved in action potential propagation, the formation of the nodes, axon–glia interactions, and demyelination diseases.


2019 ◽  
Vol 37 (3) ◽  
pp. 31
Author(s):  
Raquel Fernández González ◽  
Marcos Íñigo Pérez Pérez

The return of institutions to the main research agenda has highlighted the importance of rules in economic analysis. The New Institutional Economics has allowed a better understanding of the case studies that concern different areas of knowledge, also the one concerning the management of natural resources. In this article, the institutional analysis focuses on the maritime domain, where two large civil liability regimes for pollution coexist (OPA 90-IMO), each in a different geographical area (United States - Europe). Therefore, a comparative analysis is made between the two large regimes of civil responsibility assignment applying them to the Prestige catastrophe. In this way, the allocation and distribution of responsibilities in the investigation and subsequent judicial process of the Prestige is compared with an alternative scenario in which the applicable compensation instruments are governed by the provisions of the Oil Polution Act of 1990 (OPA 90), in order to establish a rigorous analysis on the effects that the different norms can have in the same scenario. In the comparative established in the case of the Prestige, where the responsibilities were solved very slowly in a judicial process with high transaction costs, the application of rules governed by the OPA 90 would not count with such a high degree of imperfection. This is so, since by applying the preponderance of the evidence existing in OPA 90 there would be no mitigation for the presumed culprits. On the other hand, the agents involved in the sinking would not be limited only to the owner, but also that operators or shipowners would be responsible as well. In addition, the amount of compensation would increase when counting in the damage count the personal damages, the taxes without perceiving and the ecological damage caused in a broad sense, damages not computable in the IMO.


2021 ◽  
Vol 22 (12) ◽  
pp. 6401
Author(s):  
Younglan Lim ◽  
Nam-On Ku

Although hepatocellular carcinoma (HCC) is developed with various etiologies, protection of hepatocytes seems basically essential to prevent the incidence of HCC. Keratin 8 and keratin 18 (K8/K18) are cytoskeletal intermediate filament proteins that are expressed in hepatocytes. They maintain the cell shape and protect cells under stress conditions. Their protective roles in liver damage have been described in studies of mouse models, and K8/K18 mutation frequency in liver patients. Interestingly, K8/K18 bind to signaling proteins such as transcription factors and protein kinases involved in HCC development. Since K8/K18 are abundant cytoskeletal proteins, K8/K18 binding with the signaling factors can alter the availability of the factors. Herein, we discuss the potential roles of K8/K18 in HCC development.


Open Biology ◽  
2015 ◽  
Vol 5 (8) ◽  
pp. 150080 ◽  
Author(s):  
Catherine E. Au ◽  
Louis Hermo ◽  
Elliot Byrne ◽  
Jeffrey Smirle ◽  
Ali Fazel ◽  
...  

Discovered in 1909 by Retzius and described mainly by morphology, the cytoplasmic droplet of sperm (renamed here the Hermes body) is conserved among all mammalian species but largely undefined at the molecular level. Tandem mass spectrometry of the isolated Hermes body from rat epididymal sperm characterized 1511 proteins, 43 of which were localized to the structure in situ by light microscopy and two by quantitative electron microscopy localization. Glucose transporter 3 (GLUT-3) glycolytic enzymes, selected membrane traffic and cytoskeletal proteins were highly abundant and concentrated in the Hermes body. By electron microscope gold antibody labelling, the Golgi trafficking protein TMED7/p27 localized to unstacked flattened cisternae of the Hermes body, as did GLUT-3, the most abundant protein. Its biogenesis was deduced through the mapping of protein expression for all 43 proteins during male germ cell differentiation in the testis. It is at the terminal step 19 of spermiogenesis that the 43 characteristic proteins accumulated in the nascent Hermes body.


2001 ◽  
Vol 85 (6) ◽  
pp. 2350-2358 ◽  
Author(s):  
Sanjiv K. Talwar ◽  
Pawel G. Musial ◽  
George L. Gerstein

Studies in several mammalian species have demonstrated that bilateral ablations of the auditory cortex have little effect on simple sound intensity and frequency-based behaviors. In the rat, for example, early experiments have shown that auditory ablations result in virtually no effect on the rat's ability to either detect tones or discriminate frequencies. Such lesion experiments, however, typically examine an animal's performance some time after recovery from ablation surgery. As such, they demonstrate that the cortex is not essential for simple auditory behaviors in the long run. Our study further explores the role of cortex in basic auditory perception by examining whether the cortex is normally involved in these behaviors. In these experiments we reversibly inactivated the rat primary auditory cortex (AI) using the GABA agonist muscimol, while the animals performed a simple auditory task. At the same time we monitored the rat's auditory activity by recording auditory evoked potentials (AEP) from the cortical surface. In contrast to lesion studies, the rapid time course of these experimental conditions preclude reorganization of the auditory system that might otherwise compensate for the loss of cortical processing. Soon after bilateral muscimol application to their AI region, our rats exhibited an acute and profound inability to detect tones. After a few hours this state was followed by a gradual recovery of normal hearing, first of tone detection and, much later, of the ability to discriminate frequencies. Surface muscimol application, at the same time, drastically altered the normal rat AEP. Some of the normal AEP components vanished nearly instantaneously to unveil an underlying waveform, whose size was related to the severity of accompanying behavioral deficits. These results strongly suggest that the cortex is directly involved in basic acoustic processing. Along with observations from accompanying multiunit experiments that related the AEP to AI neuronal activity, our results suggest that a critical amount of activity in the auditory cortex is necessary for normal hearing. It is likely that the involvement of the cortex in simple auditory perceptions has hitherto not been clearly understood because of underlying recovery processes that, in the long-term, safeguard fundamental auditory abilities after cortical injury.


2020 ◽  
Vol 10 (5) ◽  
pp. 6187-6190
Author(s):  
A. S. Alshammari

The keyspace of a cryptography system must be long enough in order to protect it from brute force attacks. The One-Time Pad (OTP) encryption is unconditionally secure because of its truly random keystream that is used only once. This paper proposes a new chaotic symmetric cryptosystem approach, comparable to OTP. The proposed system utilizes two Lorenz generators, a main and an auxiliary, where the aim of the second one is to make one of the main Lorenz generator’s parameters to vary continually with time in a chaotic manner. This technique was built on digitizing two Lorenz chaotic models to increase the security level. The scrambling scheme was developed and the Lorenz stream cipher binary stream successfully passed the NIST randomness test. The cryptosystem showed a high degree of security, as it had a keyspace of 2576, and it was compared with existing symmetric key cryptography systems, such as DES, 3DES, AES, Blowfish, and OTP.


Author(s):  
Weikang Qian ◽  
John Backes ◽  
Marc D. Riedel

Emerging technologies for nanoscale computation such as self-assembled nanowire arrays present specific challenges for logic synthesis. On the one hand, they provide an unprecedented density of bits with a high degree of parallelism. On the other hand, they are characterized by high defect rates. Also they often exhibit inherent randomness in the interconnects due to the stochastic nature of self-assembly. We describe a general method for synthesizing logic that exploits both the parallelism and the random effects. Our approach is based on stochastic computation with parallel bit streams. Circuits are synthesized through functional decomposition with symbolic data structures called multiplicative binary moment diagrams. Synthesis produces designs with randomized parallel components—and operations and multiplexing—that are readily implemented in nanowire crossbar arrays. Synthesis results for benchmarks circuits show that our technique maps circuit designs onto nanowire arrays effectively.


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