scholarly journals Novel roles for well-known players: from tobacco mosaic virus pests to enzymatically active assemblies

2016 ◽  
Vol 7 ◽  
pp. 613-629 ◽  
Author(s):  
Claudia Koch ◽  
Fabian J Eber ◽  
Carlos Azucena ◽  
Alexander Förste ◽  
Stefan Walheim ◽  
...  
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Self Assembly ◽  
Building Blocks ◽  
Mosaic Disease ◽  
Inorganic Materials ◽  
High Yield ◽  
External Control ◽  
Immobilization Of Enzymes

The rod-shaped nanoparticles of the widespread plant pathogentobacco mosaic virus(TMV) have been a matter of intense debates and cutting-edge research for more than a hundred years. During the late 19th century, their behavior in filtration tests applied to the agent causing the 'plant mosaic disease' eventually led to the discrimination of viruses from bacteria. Thereafter, they promoted the development of biophysical cornerstone techniques such as electron microscopy and ultracentrifugation. Since the 1950s, the robust, helically arranged nucleoprotein complexes consisting of a single RNA and more than 2100 identical coat protein subunits have enabled molecular studies which have pioneered the understanding of viral replication and self-assembly, and elucidated major aspects of virus–host interplay, which can lead to agronomically relevant diseases. However, during the last decades, TMV has acquired a new reputation as a well-defined high-yield nanotemplate with multivalent protein surfaces, allowing for an ordered high-density presentation of multiple active molecules or synthetic compounds. Amino acid side chains exposed on the viral coat may be tailored genetically or biochemically to meet the demands for selective conjugation reactions, or to directly engineer novel functionality on TMV-derived nanosticks. The natural TMV size (length: 300 nm) in combination with functional ligands such as peptides, enzymes, dyes, drugs or inorganic materials is advantageous for applications ranging from biomedical imaging and therapy approaches over surface enlargement of battery electrodes to the immobilization of enzymes. TMV building blocks are also amenable to external control of in vitro assembly and re-organization into technically expedient new shapes or arrays, which bears a unique potential for the development of 'smart' functional 3D structures. Among those, materials designed for enzyme-based biodetection layouts, which are routinely applied, e.g., for monitoring blood sugar concentrations, might profit particularly from the presence of TMV rods: Their surfaces were recently shown to stabilize enzymatic activities upon repeated consecutive uses and over several weeks. This review gives the reader a ride through strikingly diverse achievements obtained with TMV-based particles, compares them to the progress with related viruses, and focuses on latest results revealing special advantages for enzyme-based biosensing formats, which might be of high interest for diagnostics employing 'systems-on-a-chip'.

scholarly journals Milestones in research on tobacco mosaic virus

1999 ◽  
Vol 354 (1383) ◽  
pp. 521-529 ◽  
Author(s):  
B. D. Harrison ◽  
T. M. A. Wilson
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Virus Resistance ◽  
Dominant Role ◽  
Biological Properties ◽  
Mosaic Disease ◽  
Genetically Engineered ◽  
High Yield ◽  
Past Century ◽  
Avirulence Genes

Beijerinck's (1898) recognition that the cause of tobacco mosaic disease was a novel kind of pathogen became the breakthrough which led eventually to the establishment of virology as a science. Research on this agent, tobacco mosaic virus (TMV), has continued to be at the forefront of virology for the past century. After an initial phase, in which numerous biological properties of TMV were discovered, its particles were the first shown to consist of RNA and protein, and X–ray diffraction analysis of their structure was the first of a helical nucleoprotein. In the molecular biological phase of research, TMV RNA was the first plant virus genome to be sequenced completely, its genes were found to be expressed by cotranslational particle disassembly and the use of subgenomic mRNA, and the mechanism of assembly of progeny particles from their separate parts was discovered. Molecular genetical and cell biological techniques were then used to clarify the roles and modes of action of the TMV non–structural proteins: the 126 kDa and 183 kDa replicase components and the 30 kDa cell–to–cell movement protein. Three different TMV genes were found to act as avirulence genes, eliciting hypersensitive responses controlled by specific, but different, plant genes. One of these (the N gene) was the first plant gene controlling virus resistance to be isolated and sequenced. In the biotechnological sphere, TMV has found several applications: as the first source of transgene sequences conferring virus resistance, in vaccines consisting of TMV particles genetically engineered to carry foreign epitopes, and in systems for expressing foreign genes. TMV owes much of its popularity as a research model to the great stability and high yield of its particles. Although modern methods have much decreased the need for such properties, and TMV may have a less dominant role in the future, it continues to occupy a prominent position in both fundamental and applied research.

scholarly journals Bottom-Up Fabrication of Protein Nanowires via Controlled Self-Assembly of Recombinant Geobacter Pilins

mBio ◽  
2019 ◽  
Vol 10 (6) ◽  
Author(s):  
K. M. Cosert ◽  
Angelines Castro-Forero ◽  
Rebecca J. Steidl ◽  
Robert M. Worden ◽  
G. Reguera
Keyword(s):  
Electronic Properties ◽  
Self Assembly ◽  
Building Blocks ◽  
Geobacter Sulfurreducens ◽  
Helical Conformation ◽  
High Yield ◽  
Efficient Production ◽  
Bottom Up ◽  
Content Type

ABSTRACT Metal-reducing bacteria in the genus Geobacter use a complex protein apparatus to guide the self-assembly of a divergent type IVa pilin peptide and synthesize conductive pilus appendages that show promise for the sustainable manufacturing of protein nanowires. The preferential helical conformation of the Geobacter pilin, its high hydrophobicity, and precise distribution of charged and aromatic amino acids are critical for biological self-assembly and conductivity. We applied this knowledge to synthesize via recombinant methods truncated pilin peptides for the bottom-up fabrication of protein nanowires and identified rate-limiting steps of pilin nucleation and fiber elongation that control assembly efficiency and nanowire length, respectively. The synthetic fibers retained the biochemical and electronic properties of the native pili even under chemical fixation, a critical consideration for integration of the nanowires into electronic devices. The implications of these results for the design and mass production of customized protein nanowires for diverse applications are discussed. IMPORTANCE The discovery in 2005 of conductive protein appendages (pili) in the metal-reducing bacterium Geobacter sulfurreducens challenged our understanding of biological electron transfer and pioneered studies in electromicrobiology that revealed the electronic basis of many microbial metabolisms and interactions. The protein nature of the pili afforded opportunities for engineering novel conductive peptides for the synthesis of nanowires via cost-effective and scalable manufacturing approaches. However, methods did not exist for efficient production, purification, and in vitro assembly of pilins into nanowires. Here we describe platforms for high-yield recombinant synthesis of Geobacter pilin derivatives and their assembly as protein nanowires with biochemical and electronic properties rivaling those of the native pili. The bottom-up fabrication of protein nanowires exclusively from pilin building blocks confirms unequivocally the charge transport capacity of the peptide assembly and establishes the intellectual foundation needed to manufacture pilin-based nanowires in bioelectronics and other applications.

scholarly journals Self–assembly of tobacco mosaic virus: the role of an intermediate aggregate in generating both specificity and speed

1999 ◽  
Vol 354 (1383) ◽  
pp. 537-550 ◽  
Author(s):  
P. J. G. Butler
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Self Assembly ◽  
Specific Binding ◽  
Macromolecular Structure ◽  
Stem Loop ◽  
Protein Subunits ◽  
Internal Site

The tobacco mosaic virus (TMV) particle was the first macromolecular structure to be shown to self–assemble in vitro , allowing detailed studies of the mechanism. Nucleation of TMV self–assembly is by the binding of a specific stem–loop of the single–stranded viral RNA into the central hole of a two–ring sub–assembly of the coat protein, known as the ‘disk’. Binding of the loop onto its specific binding site, between the two rings of the disk, leads to melting of the stem so more RNA is available to bind. The interaction of the RNA with the protein subunits in the disk cause this to dislocate into a proto–helix, rearranging the protein subunits in such a way that the axial gap between the rings at inner radii closes, entrapping the RNA. Assembly starts at an internal site on TMV RNA, about 1 kb from its 3′–terminus, and the elongation in the two directions is different. Elongation of the nucleated rods towards the 5′–terminus occurs on a ‘travelling loop’ of the RNA and, predominantly, still uses the disk sub–assembly of protein subunits, consequently incorporating approximately 100 further nucleotides as each disk is added, while elongation towards the 3′–terminus uses smaller protein aggregates and does not show this ’quantized‘ incorporation.

scholarly journals Constructing Large 2D Lattices Out of DNA-Tiles

Molecules ◽  
2021 ◽  
Vol 26 (6) ◽  
pp. 1502
Author(s):  
Johannes M. Parikka ◽  
Karolina Sokołowska ◽  
Nemanja Markešević ◽  
J. Jussi Toppari
Keyword(s):  
Self Assembly ◽  
Dna Nanotechnology ◽  
Building Blocks ◽  
High Yield ◽  
Dna Nanostructures ◽  
Liquid Interfaces ◽  
Basic Principles ◽  
Dna Tiles ◽  
One Step ◽  
Solid Liquid

The predictable nature of deoxyribonucleic acid (DNA) interactions enables assembly of DNA into almost any arbitrary shape with programmable features of nanometer precision. The recent progress of DNA nanotechnology has allowed production of an even wider gamut of possible shapes with high-yield and error-free assembly processes. Most of these structures are, however, limited in size to a nanometer scale. To overcome this limitation, a plethora of studies has been carried out to form larger structures using DNA assemblies as building blocks or tiles. Therefore, DNA tiles have become one of the most widely used building blocks for engineering large, intricate structures with nanometer precision. To create even larger assemblies with highly organized patterns, scientists have developed a variety of structural design principles and assembly methods. This review first summarizes currently available DNA tile toolboxes and the basic principles of lattice formation and hierarchical self-assembly using DNA tiles. Special emphasis is given to the forces involved in the assembly process in liquid-liquid and at solid-liquid interfaces, and how to master them to reach the optimum balance between the involved interactions for successful self-assembly. In addition, we focus on the recent approaches that have shown great potential for the controlled immobilization and positioning of DNA nanostructures on different surfaces. The ability to position DNA objects in a controllable manner on technologically relevant surfaces is one step forward towards the integration of DNA-based materials into nanoelectronic and sensor devices.

scholarly journals Role of the 3′ tRNA-Like Structure in Tobacco Mosaic Virus Minus-Strand RNA Synthesis by the Viral RNA-Dependent RNA Polymerase In Vitro

2000 ◽  
Vol 74 (24) ◽  
pp. 11671-11680 ◽  
Author(s):  
T. A. M. Osman ◽  
C. L. Hemenway ◽  
K. W. Buck
Keyword(s):  
Rna Polymerase ◽  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Untranslated Region ◽  
Rna Synthesis ◽  
Central Core ◽  
Minus Strand ◽  
Stem Loop ◽  
Polymerase Binding

ABSTRACT A template-dependent RNA polymerase has been used to determine the sequence elements in the 3′ untranslated region of tobacco mosaic virus RNA that are required for promotion of minus-strand RNA synthesis and binding to the RNA polymerase in vitro. Regions which were important for minus-strand synthesis were domain D1, which is equivalent to a tRNA acceptor arm; domain D2, which is similar to a tRNA anticodon arm; an upstream domain, D3; and a central core, C, which connects domains D1, D2, and D3 and determines their relative orientations. Mutational analysis of the 3′-terminal 4 nucleotides of domain D1 indicated the importance of the 3′-terminal CA sequence for minus-strand synthesis, with the sequence CCCA or GGCA giving the highest transcriptional efficiency. Several double-helical regions, but not their sequences, which are essential for forming pseudoknot and/or stem-loop structures in domains D1, D2, and D3 and the central core, C, were shown to be required for high template efficiency. Also important were a bulge sequence in the D2 stem-loop and, to a lesser extent, a loop sequence in a hairpin structure in domain D1. The sequence of the 3′ untranslated region upstream of domain D3 was not required for minus-strand synthesis. Template-RNA polymerase binding competition experiments showed that the highest-affinity RNA polymerase binding element region lay within a region comprising domain D2 and the central core, C, but domains D1 and D3 also bound to the RNA polymerase with lower affinity.

An analysis of tobacco mosaic virus replicative structures synthesized in vitro

1986 ◽  
Vol 6 (6) ◽  
pp. 455-465 ◽  
Author(s):  
Nevin Dale Young ◽  
Milton Zaitlin
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus

scholarly journals Tobacco Mosaic Virus Movement Protein Functions as a Structural Microtubule-Associated Protein

2006 ◽  
Vol 80 (17) ◽  
pp. 8329-8344 ◽  
Author(s):  
Jamie Ashby ◽  
Emmanuel Boutant ◽  
Mark Seemanpillai ◽  
Adrian Sambade ◽  
Christophe Ritzenthaler ◽  
...  
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Movement Protein ◽  
Cell Extract ◽  
Transport Processes ◽  
In Vitro Assays ◽  
Protein Functions ◽  
Intercellular Spread

ABSTRACT The cell-to-cell spread of Tobacco mosaic virus infection depends on virus-encoded movement protein (MP), which is believed to form a ribonucleoprotein complex with viral RNA (vRNA) and to participate in the intercellular spread of infectious particles through plasmodesmata. Previous studies in our laboratory have provided evidence that the vRNA movement process is correlated with the ability of the MP to interact with microtubules, although the exact role of this interaction during infection is not known. Here, we have used a variety of in vivo and in vitro assays to determine that the MP functions as a genuine microtubule-associated protein that binds microtubules directly and modulates microtubule stability. We demonstrate that, unlike MP in whole-cell extract, microtubule-associated MP is not ubiquitinated, which strongly argues against the hypothesis that microtubules target the MP for degradation. In addition, we found that MP interferes with kinesin motor activity in vitro, suggesting that microtubule-associated MP may interfere with kinesin-driven transport processes during infection.

In vitro and in vivo aggregation of the defective PM2 tobacco mosaic virus protein

1966 ◽  
Vol 19 (1) ◽  
pp. 140-IN8 ◽  
Author(s):  
Albert Siegel ◽  
G.J. Hills ◽  
Roy Markham
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Virus Protein

On in vitro and in vivo binding of tobacco mosaic virus to cytoplasmic membranes

1984 ◽  
Vol 179 (6) ◽  
pp. 507-516 ◽  
Author(s):  
Barbara Pustowoit ◽  
Wladimir Pustowoit ◽  
Gottfried Schuster
Keyword(s):  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
In Vivo Binding ◽  
Cytoplasmic Membranes

High Yield Assembly of Compliant MEMS Snap Fasteners

2008 ◽  
Author(s):  
Rakesh Murthy ◽  
Aditya N. Das ◽  
Dan O. Popa
Keyword(s):  
Self Assembly ◽  
Building Blocks ◽  
Deformation Energy ◽  
High Yield ◽  
3 Dimensional ◽  
Lab Experiments ◽  
Trade Offs ◽  
Robotic Cells ◽  
And Performance ◽  
Important Design

Heterogeneous assembly at the microscale has recently emerged as a viable pathway to constructing 3-dimensional microrobots and other miniaturized devices. In contrast to self-assembly, this method is directed and deterministic, and is based on serial or parallel microassembly. Whereas at the meso and macro scales, automation is often undertaken after, and often benchmarked against manual assembly, we demonstrate that deterministic automation at the MEMS scale can be completed with higher yields through the use of engineered compliance and precision robotic cells. Snap fasteners have long been used as a way to exploit the inherent stability of local minima of the deformation energy caused by interference during part mating. In this paper we assume that the building blocks are 2 1/2 -dimensional, as is the case with lithographically microfabricated MEMS parts. The assembly of the snap fasteners is done using μ3, a multi-robot microassembly station with unique characteristics located at our ARRI’s Texas Microfactory lab. Experiments are performed to demonstrate that fast and reliable assemblies can be expected if the microparts and the robotic cell satisfy a so-called “High Yield Assembly Condition” (H.Y.A.C.). Important design trade-offs for assembly and performance of microsnap fasteners are discussed and experimentally evaluated.

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