Directed Therapies to Nucleic Acid and Cytoplasmic RNA

2021 ◽  
pp. 445-458
Author(s):  
Mitali Ghose
Keyword(s):  
Nucleic Acid ◽  
Cytoplasmic Rna

scholarly journals Cytological and Biochemical Studies of Anucleolate Xenopus Larvae

1962 ◽  
Vol s3-103 (61) ◽  
pp. 25-35
Author(s):  
H. WALLACE
Keyword(s):  
Alkaline Phosphatase ◽  
Nucleic Acid ◽  
Acid Content ◽  
Deoxyribonucleic Acid ◽  
Nucleic Acid Content ◽  
Tail Bud ◽  
Lethal Mutant ◽  
Cytoplasmic Rna ◽  
Biochemical Studies ◽  
And Function

A larval lethal mutant of Xenopus laevis lacks true nucleoli but possesses analogous intranuclear organelles, here termed blobs, which are smaller and more numerous than nucleoli. Cytochemical tests reveal that blobs (like nucleoli) contain ribonucleic acid (RNA), arginine, and alkaline phosphatase, but probably no Feulgen-positive material. Anucleolate larvae are deficient in cytoplasmic RNA. By biochemical methods the nucleic acid content of anucleolate embryos is found to be normal at the tail-bud stage, but does not increase after this. By the time they hatch, anucleolate larvae are deficient in both RNA and deoxyribonucleic acid (DNA). The implications of this and related mutations on the formation and function of the nucleolus are considered. The term ‘blob’ is justified in that it would be misleading to regard such organelles as nucleoli produced by normally latent organizers.

scholarly journals Methyl-Green-Pyronin as a Differential Nucleic Acid Stain for Peripheral Blood Smears

Blood ◽  
1956 ◽  
Vol 11 (12) ◽  
pp. 1132-1139 ◽  
Author(s):  
SEYMOUR PERRY ◽  
JOHN REYNOLDS ◽  
MARY BAKER
Keyword(s):  
Nucleic Acid ◽  
Pernicious Anemia ◽  
Peripheral Blood ◽  
Blood Cells ◽  
White Blood Cells ◽  
Cell Physiology ◽  
Methyl Green ◽  
Blood Leukocytes ◽  
Cytoplasmic Rna ◽  
Nucleic Acid Stain

Abstract A differential nucleic acid stain for peripheral blood and marrow smears using methyl-green and pyronin Y and Carnoy’s solution as a fixative is described. Characteristics of the various blood cells are quite distinct so that identification is readily accomplished. If pyroninophilia is assumed to be a measure of RNA, morphologically identical and mature leukocytes are found to vary in the quantity of cytoplasmic RNA. Since cellular immaturity is associated with increased RNA, this must be an indication of the varying "age" of white blood cells in the peripheral blood. With the use of this stain a much more dynamic picture of white cell physiology is obtained than with other more conventional stains. The quantity of cytoplasmic RNA in peripheral blood leukocytes, as measured by a scoring technic, was found to be increased as compared to normal in most infections, leukemias, pernicious anemia in relapse, and a variety of other conditions. With treatment, the score fell to normal levels in pernicious anemia and infections.

scholarly journals Unique spectral signatures of the nucleic acid dye acridine orange can distinguish cell death by apoptosis and necroptosis

2017 ◽  
Vol 216 (4) ◽  
pp. 1163-1181 ◽  
Author(s):  
Jason R. Plemel ◽  
Andrew V. Caprariello ◽  
Michael B. Keough ◽  
Tyler J. Henry ◽  
Shigeki Tsutsui ◽  
...  
Keyword(s):  
Cell Death ◽  
Nucleic Acid ◽  
Acridine Orange ◽  
Apoptotic Cell ◽  
Wide Spectrum ◽  
Experimental Models ◽  
Cellular Injury ◽  
Acid Dye ◽  
Spectral Signatures ◽  
Cytoplasmic Rna

Cellular injury and death are ubiquitous features of disease, yet tools to detect them are limited and insensitive to subtle pathological changes. Acridine orange (AO), a nucleic acid dye with unique spectral properties, enables real-time measurement of RNA and DNA as proxies for cell viability during exposure to various noxious stimuli. This tool illuminates spectral signatures unique to various modes of cell death, such as cells undergoing apoptosis versus necrosis/necroptosis. This new approach also shows that cellular RNA decreases during necrotic, necroptotic, and apoptotic cell death caused by demyelinating, ischemic, and traumatic injuries, implying its involvement in a wide spectrum of tissue pathologies. Furthermore, cells with pathologically low levels of cytoplasmic RNA are detected earlier and in higher numbers than with standard markers including TdT-mediated dUTP biotin nick-end labeling and cleaved caspase 3 immunofluorescence. Our technique highlights AO-labeled cytoplasmic RNA as an important early marker of cellular injury and a sensitive indicator of various modes of cell death in a range of experimental models.

scholarly journals Regulation of signaling mediated by nucleic acid sensors for innate interferon-mediated responses during viral infection

2019 ◽  
Vol 31 (8) ◽  
pp. 477-488 ◽  
Author(s):  
Akinori Takaoka ◽  
Taisho Yamada
Keyword(s):  
Nucleic Acid ◽  
Viral Infection ◽  
Type I ◽  
Post Translational Modification ◽  
Dna Viruses ◽  
Downstream Signaling ◽  
New Class ◽  
Cytoplasmic Rna ◽  
Sensor Activation ◽  
Acid Sensors

Abstract Type I and type III interferons are important anti-viral cytokines that are massively induced during viral infection. This dynamic process is regulated by many executors and regulators for efficient eradication of invading viruses and protection from harmful, excessive responses. An array of innate sensors recognizes virus-derived nucleic acids to activate their downstream signaling to evoke cytokine responses including interferons. In particular, a cytoplasmic RNA sensor RIG-I (retinoic acid-inducible gene I) is involved in the detection of multiple types of not only RNA viruses but also DNA viruses. Accumulating findings have revealed that activation of nucleic acid sensors and the related signaling mediators is regulated on the basis of post-translational modification such as ubiquitination, phosphorylation and ADP-ribosylation. In addition, long non-coding RNAs (lncRNAs) have been implicated as a new class of regulators in innate signaling. A comprehensive understanding of the regulatory mechanisms of innate sensor activation and its signaling in host–virus interaction will provide a better therapeutic strategy to efficiently control viral infection and maintain immune homeostasis.

Introduction

1978 ◽  
Vol 36 (3) ◽  
pp. 543-544
Author(s):  
W. Bernard
Keyword(s):  
Molecular Weight ◽  
Electron Microscope ◽  
Nucleic Acid ◽  
Gene Mapping ◽  
Molecular Genetics ◽  
Cell Biology ◽  
Gene Activity ◽  
Close Connection ◽  
Basic Information ◽  
Simple Systems

In comparison to many other fields of ultrastructural research in Cell Biology, the successful exploration of genes and gene activity with the electron microscope in higher organisms is a late conquest. Nucleic acid molecules of Prokaryotes could be successfully visualized already since the early sixties, thanks to the Kleinschmidt spreading technique - and much basic information was obtained concerning the shape, length, molecular weight of viral, mitochondrial and chloroplast nucleic acid. Later, additonal methods revealed denaturation profiles, distinction between single and double strandedness and the use of heteroduplexes-led to gene mapping of relatively simple systems carried out in close connection with other methods of molecular genetics.

Infection of Escherichia coli with bacteriophages

1969 ◽  
Vol 27 ◽  
pp. 370-371
Author(s):  
Manfred E. Bayer
Keyword(s):  
Escherichia Coli ◽  
Nucleic Acid ◽  
Cell Surface ◽  
Virus Type ◽  
Infection Process ◽  
Adsorption Site ◽  
The Other ◽  
Specific Receptors ◽  
Bacterial Receptors

The first step in the infection of a bacterium by a virus consists of a collision between cell and bacteriophage. The presence of virus-specific receptors on the cell surface will trigger a number of events leading eventually to release of the phage nucleic acid. The execution of the various "steps" in the infection process varies from one virus-type to the other, depending on the anatomy of the virus. Small viruses like ØX 174 and MS2 adsorb directly with their capsid to the bacterial receptors, while other phages possess attachment organelles of varying complexity. In bacteriophages T3 (Fig. 1) and T7 the small conical processes of their heads point toward the adsorption site; a welldefined baseplate is attached to the head of P22; heads without baseplates are not infective.

Nucleic Acid Molecules Prepared by Monolayer Techniques

1972 ◽  
Vol 30 ◽  
pp. 178-179
Author(s):  
Dimitrij Lang
Keyword(s):  
Electron Microscopy ◽  
Standard Deviation ◽  
Nucleic Acid ◽  
Cytochrome C ◽  
Nucleic Acids ◽  
Contour Length ◽  
Sample Standard Deviation ◽  
Molecular Weights ◽  
Length Measurements ◽  
Protein Monolayer

The success of the protein monolayer technique for electron microscopy of individual DNA molecules is based on the prevention of aggregation and orientation of the molecules during drying on specimen grids. DNA adsorbs first to a surface-denatured, insoluble cytochrome c monolayer which is then transferred to grids, without major distortion, by touching. Fig. 1 shows three basic procedures which, modified or not, permit the study of various important properties of nucleic acids, either in concert with other methods or exclusively:1) Molecular weights relative to DNA standards as well as number distributions of molecular weights can be obtained from contour length measurements with a sample standard deviation between 1 and 4%.

Snapshot blotting: The transfer of nucleic acids and nucleoprotein complexes from electrophoresis gels to EM grids

1992 ◽  
Vol 50 (1) ◽  
pp. 762-763
Author(s):  
Stephen D. Jett
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Nucleic Acid Binding ◽  
Mobility Shift ◽  
Carbon Coated ◽  
Nucleoprotein Complexes ◽  
Mobility Shift Assay ◽  
Protein Nucleic Acid ◽  
Gel Mobility Shift ◽  
Nucleic Acid Interactions

The electrophoresis gel mobility shift assay is a popular method for the study of protein-nucleic acid interactions. The binding of proteins to DNA is characterized by a reduction in the electrophoretic mobility of the nucleic acid. Binding affinity, stoichiometry, and kinetics can be obtained from such assays; however, it is often desirable to image the various species in the gel bands using TEM. Present methods for isolation of nucleoproteins from gel bands are inefficient and often destroy the native structure of the complexes. We have developed a technique, called “snapshot blotting,” by which nucleic acids and nucleoprotein complexes in electrophoresis gels can be electrophoretically transferred directly onto carbon-coated grids for TEM imaging.

High-resolution nucleic acid sequence mapping via in situ hybridization at the Electron Microscope level

1995 ◽  
Vol 53 ◽  
pp. 752-753
Author(s):  
B.A. Hamkalo ◽  
S. Narayanswami ◽  
A.P. Kausch
Keyword(s):  
Nucleic Acid ◽  
Interphase Nucleus ◽  
Genome Mapping ◽  
Precise Location ◽  
Electron Microscope Level ◽  
Rna Sequences ◽  
Fundamental Interest ◽  
Whole Cells ◽  
Dna And Rna

The availability of nonradioactive methods to label nucleic acids an the resultant rapid and greater sensitivity of detection has catapulted the technique of in situ hybridization to become the method of choice to locate of specific DNA and RNA sequences on chromosomes and in whole cells in cytological preparations in many areas of biology. It is being applied to problems of fundamental interest to basic cell and molecular biologists such as the organization of the interphase nucleus in the context of putative functional domains; it is making major contributions to genome mapping efforts; and it is being applied to the analysis of clinical specimens. Although fluorescence detection of nucleic acid hybrids is routinely used, certain questions require greater resolution. For example, very closely linked sequences may not be separable using fluorescence; the precise location of sequences with respect to chromosome structures may be below the resolution of light microscopy(LM); and the relative positions of sequences on very small chromosomes may not be feasible.

Imaging of ribosomal RNA-protein interactions by dark-field STEM

1989 ◽  
Vol 47 ◽  
pp. 250-251
Author(s):  
M. Boublik ◽  
V. Mandiyan ◽  
S. Tumminia ◽  
J.F. Hainfeld ◽  
J.S. Wall
Keyword(s):  
Nucleic Acid ◽  
16S Rrna ◽  
Dark Field ◽  
Radius Of Gyration ◽  
Central Domain ◽  
The Core ◽  
16S Rna ◽  
30S Subunit ◽  
Core Particles

Success in protein-free deposition of native nucleic acid molecules from solutions of selected ionic conditions prompted attempts for high resolution imaging of nucleic acid interactions with proteins, not attainable by conventional EM. Since the nucleic acid molecules can be visualized in the dark-field STEM mode without contrasting by heavy atoms, the established linearity between scattering cross-section and molecular weight can be applied to the determination of their molecular mass (M) linear density (M/L), mass distribution and radius of gyration (RG). Determination of these parameters promotes electron microscopic imaging of biological macromolecules by STEM to a quantitative analytical level. This technique is applied to study the mechanism of 16S rRNA folding during the assembly process of the 30S ribosomal subunit of E. coli. The sequential addition of protein S4 which binds to the 5'end of the 16S rRNA and S8 and S15 which bind to the central domain of the molecule leads to a corresponding increase of mass and increased coiling of the 16S rRNA in the core particles. This increased compactness is evident from the decrease in RG values from 114Å to 91Å (in “ribosomal” buffer consisting of 10 mM Hepes pH 7.6, 60 mM KCl, 2 m Mg(OAc)2, 1 mM DTT). The binding of S20, S17 and S7 which interact with the 5'domain, the central domain and the 3'domain, respectively, continues the trend of mass increase. However, the RG values of the core particles exhibit a reverse trend, an increase to 108Å. In addition, the binding of S7 leads to the formation of a globular mass cluster with a diameter of about 115Å and a mass of ∽300 kDa. The rest of the mass, about 330 kDa, remains loosely coiled giving the particle a “medusa-like” appearance. These results provide direct evidence that 16S RNA undergoes significant structural reorganization during the 30S subunit assembly and show that its interactions with the six primary binding proteins are not sufficient for 16S rRNA coiling into particles resembling the native 30S subunit, contrary to what has been reported in the literature.

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