scholarly journals A LABILE INTRANUCLEAR RNA ASSOCIATED WITH THE DEVELOPMENT OF ADENOVIRUSES

1964 ◽  
Vol 119 (3) ◽  
pp. 433-442 ◽  
Author(s):  
Heather Donald Mayor
Keyword(s):  
Nucleic Acids ◽  
Acridine Orange ◽  
Viral Protein ◽  
Rna Viruses ◽  
Monkey Kidney ◽  
Kidney Cells ◽  
Viral Dna ◽  
Dna And Rna ◽  
Double Stranded Dna ◽  
Cellular Dna

The nucleic acids produced intracellularly during the replication cycles of both DNA and RNA viruses can now be identified rapidly using a sensitized procedure based on staining with the fluorochrome acridine orange. Cellular DNA, viral DNA (both single and double stranded forms), cellular RNA, and RNA arising as a result of viral stimulus can be differentiated. The intracellular development of virus specific DNA, RNA, and protein has been studied in monkey kidney cells infected with adenoviruses types 3 and 7. It has been possible to detect a labile RNA in the nucleus from 16 to 20 hours after inoculation. When the cultures are treated with puromycin at this time, this RNA can be accumulated under certain conditions in the nucleus and demonstrated cytochemically. At the same time the production of specific viral protein as determined by staining with fluorescein-labeled antibodies is markedly inhibited. However, intranuclear double stranded DNA continues to be formed for a time. When puromycin is added to the system early in the eclipse period virus-specific DNA and labile RNA cannot be detected.

scholarly journals CYTOLOGICAL AND CYTOCHEMICAL STUDIES OF GREEN MONKEY KIDNEY CELLS INFECTED IN VITRO WITH SIMIAN VIRUS 40

1965 ◽  
Vol 25 (3) ◽  
pp. 529-543 ◽  
Author(s):  
Robert Love ◽  
Mario V. Fernandes
Keyword(s):  
Nucleic Acids ◽  
Acridine Orange ◽  
Toluidine Blue ◽  
Trichloroacetic Acid ◽  
Simian Virus 40 ◽  
Monkey Kidney ◽  
Methyl Green ◽  
Kidney Cells ◽  
Green Monkey ◽  
Inclusion Material

Cytological and cytochemical studies of green monkey kidney cells infected with SV40 virus indicated that the type of lesion produced was influenced by the multiplicity of infection and that the lesions appeared later and progressed more slowly when the inoculum was diluted. The earliest change consisted of enlargement of ribonucleoprotein-containing spherules in the nucleolus (nucleolini). This was followed by rarefaction, with or without condensation, of the chromatin and the appearance of one or more homogeneous masses of inclusion material containing DNA, RNA, and non-histone protein which eventually filled the nucleus. In some instances the chromatin appeared to be directly transformed into inclusion material. In the later stages of infection, the ribonucleoprotein of the nucleolini was no longer stainable and material resembling the nucleoprotein of the intranuclear inclusions was found in the nucleolar vacuoles and in the cytoplasm. The nucleic acids in the inclusions were stained by toluidine blue, toluidine blue-molybdate, the Feulgen stain, and by methyl green. The stainable material was extractable by nuclease digestion or by hot trichloroacetic acid. Green or yellowish green staining by acridine orange was apparently due to binding of dye by protein and not by nucleic acids since the staining reaction was not reduced by extraction of nucleic acids by hot trichloroacetic acid. Extraction with pepsin in combination with ribonuclease or deoxyribonuclease removed practically all the inclusions from the cells; consequently they could not be stained with acridine orange. The cytochemical studies suggest that the use of pepsin together with nuclease is not a meaningful technique.

scholarly journals Packaging of Genomic RNA in Positive-Sense Single-Stranded RNA Viruses: A Complex Story

Viruses ◽  
2019 ◽  
Vol 11 (3) ◽  
pp. 253 ◽  
Author(s):  
Mauricio Comas-Garcia
Keyword(s):  
Rna Viruses ◽  
Genomic Rna ◽  
Rna Packaging ◽  
Dna And Rna ◽  
Relative Contribution ◽  
Double Stranded Dna ◽  
Positive Sense ◽  
Infectious Cycle

The packaging of genomic RNA in positive-sense single-stranded RNA viruses is a key part of the viral infectious cycle, yet this step is not fully understood. Unlike double-stranded DNA and RNA viruses, this process is coupled with nucleocapsid assembly. The specificity of RNA packaging depends on multiple factors: (i) one or more packaging signals, (ii) RNA replication, (iii) translation, (iv) viral factories, and (v) the physical properties of the RNA. The relative contribution of each of these factors to packaging specificity is different for every virus. In vitro and in vivo data show that there are different packaging mechanisms that control selective packaging of the genomic RNA during nucleocapsid assembly. The goals of this article are to explain some of the key experiments that support the contribution of these factors to packaging selectivity and to draw a general scenario that could help us move towards a better understanding of this step of the viral infectious cycle.

NUCLEIC ACIDS BASED DIAGNOSTICS FOR SALIVARY VIRUSES: A REVIEW

2021 ◽  
pp. 4168-4179
Keyword(s):  
Nucleic Acids ◽  
Viral Infection ◽  
Viral Infections ◽  
Genomic Analysis ◽  
Metagenomic Analysis ◽  
Viral Dna ◽  
Dna And Rna ◽  
Pathogenic Viruses ◽  
Current Detection ◽  
General Profile

Objectives: This review aims to find the general profile of salivary nucleic acids; to understand the current detection method of salivary viral DNA and RNA, and to understand the prospect of metagenomic analysis for salivary viral infection. Method: This review was started with searching processes with the help of PubMed, during August 2020. After screening the articles, 64 articles were obtained and used as referents. Results: Salivary nucleic acids contain viral DNA and RNA that can be used as biomarkers for virus infection. Even the salivary amount of DNA/RNA is very tiny, it is definite and associated with the viral infection. Viral DNA and viral RNA in saliva are categorized into two groups, the gland salivary viral DNA/RNA and systemic viral DNA/RNA. Most of the current PCR-based methods are single genomic analysis that can not detect multi strains community of pathogenic viruses. Multiple genomic analysis is needed to be implemented for multi strains of viral infection. Conclusions: The presence of viral nucleic acids in the saliva is associated closely with the glands salivary and the systemic viral infections. Therefore, they are useful biomarkers for the diagnosis of viral diseases. In the future, the metagenomic analysis can be applied for the multi strains of viral analysis.

Loading of Human Red Blood Cells with DNA and RNA

1976 ◽  
Vol 31 (3-4) ◽  
pp. 149-151 ◽  
Author(s):  
Dieter Auer ◽  
Gerhard Brandner
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Human Erythrocytes ◽  
Viral Dna ◽  
External Concentration ◽  
Dna And Rna ◽  
Human Red Blood Cells ◽  
Hank’S Solution

Abstract Human erythrocytes were suspended in Hank’s solution containing mammalian or viral DNA or RNA. After dialysis at 0 °C first against water and subsequently against Hank’s solution, and a further incubation at 37 °C , the erythrocytes were found to be loaded with the nucleic acids. The nucleic acid trapped in the erythrocytes exhibited up to 35 per cent of the external concentration.

scholarly journals Getting DNA and RNA out of the dark with 2CNqA: a bright adenine analogue and interbase FRET donor

2020 ◽  
Vol 48 (14) ◽  
pp. 7640-7652 ◽  
Author(s):  
Anna Wypijewska del Nogal ◽  
Anders F Füchtbauer ◽  
Mattias Bood ◽  
Jesper R Nilsson ◽  
Moa S Wranne ◽  
...  
Keyword(s):  
Quantum Yield ◽  
Nucleic Acids ◽  
Photophysical Properties ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Molar Absorptivity ◽  
Dna And Rna ◽  
Double Stranded Dna ◽  
Rna Duplexes

Abstract With the central role of nucleic acids there is a need for development of fluorophores that facilitate the visualization of processes involving nucleic acids without perturbing their natural properties and behaviour. Here, we incorporate a new analogue of adenine, 2CNqA, into both DNA and RNA, and evaluate its nucleobase-mimicking and internal fluorophore capacities. We find that 2CNqA displays excellent photophysical properties in both nucleic acids, is highly specific for thymine/uracil, and maintains and slightly stabilises the canonical conformations of DNA and RNA duplexes. Moreover, the 2CNqA fluorophore has a quantum yield in single-stranded and duplex DNA ranging from 10% to 44% and 22% to 32%, respectively, and a slightly lower one (average 12%) inside duplex RNA. In combination with a comparatively strong molar absorptivity for this class of compounds, the resulting brightness of 2CNqA inside double-stranded DNA is the highest reported for a fluorescent base analogue. The high, relatively sequence-independent quantum yield in duplexes makes 2CNqA promising as a nucleic acid label and as an interbase Förster resonance energy transfer (FRET) donor. Finally, we report its excellent spectral overlap with the interbase FRET acceptors qAnitro and tCnitro, and demonstrate that these FRET pairs enable conformation studies of DNA and RNA.

scholarly journals Understanding the mechanical response of double-stranded DNA and RNA under constant stretching forces using all-atom molecular dynamics

2017 ◽  
Vol 114 (27) ◽  
pp. 7049-7054 ◽  
Author(s):  
Alberto Marin-Gonzalez ◽  
J. G. Vilhena ◽  
Ruben Perez ◽  
Fernando Moreno-Herrero
Keyword(s):  
Molecular Dynamics ◽  
Nucleic Acids ◽  
Elastic Constants ◽  
Base Pair ◽  
Mechanical Response ◽  
Atomic Level ◽  
Biological Processes ◽  
Open Structure ◽  
Dna And Rna ◽  
Double Stranded Dna

Multiple biological processes involve the stretching of nucleic acids (NAs). Stretching forces induce local changes in the molecule structure, inhibiting or promoting the binding of proteins, which ultimately affects their functionality. Understanding how a force induces changes in the structure of NAs at the atomic level is a challenge. Here, we use all-atom, microsecond-long molecular dynamics to simulate the structure of dsDNA and dsRNA subjected to stretching forces up to 20 pN. We determine all of the elastic constants of dsDNA and dsRNA and provide an explanation for three striking differences in the mechanical response of these two molecules: the threefold softer stretching constant obtained for dsRNA, the opposite twist-stretch coupling, and its nontrivial force dependence. The lower dsRNA stretching resistance is linked to its more open structure, whereas the opposite twist-stretch coupling of both molecules is due to the very different evolution of molecules’ interstrand distance with the stretching force. A reduction of this distance leads to overwinding in dsDNA. In contrast, dsRNA is not able to reduce its interstrand distance and can only elongate by unwinding. Interstrand distance is directly correlated with the slide base-pair parameter and its different behavior in dsDNA and dsRNA traced down to changes in the sugar pucker angle of these NAs.

The multiple roles of the nucleocapsid in retroviral RNA conversion into proviral DNA by reverse transcriptase

2016 ◽  
Vol 44 (5) ◽  
pp. 1427-1440 ◽  
Author(s):  
Jean-Luc Darlix ◽  
Hugues de Rocquigny ◽  
Yves Mély
Keyword(s):  
Nucleic Acids ◽  
Reverse Transcriptase ◽  
Multiple Roles ◽  
Genomic Rna ◽  
Proviral Dna ◽  
Double Stranded Dna ◽  
Replication Strategy ◽  
Dimeric Form ◽  
Copy And Paste ◽  
Cellular Dna

Retroviruses are enveloped plus-strand RNA viruses that can cause cancer, immunodeficiency and neurological disorder in human and animals. Retroviruses have several unique properties, such as a genomic RNA in a dimeric form found in the virus, and a replication strategy called ‘copy-and-paste' during which the plus-strand genomic RNA is converted into a double-stranded DNA, subsequently integrated into the cellular genome. Two essential viral enzymes, reverse transcriptase (RT) and integrase (IN), direct this ‘copy-and-paste' replication. RT copies the genomic RNA generating the double-stranded proviral DNA, while IN catalyzes proviral DNA integration into the cellular DNA, then called the provirus. In that context, a major component of the virion core, the nucleocapsid protein (NC), was found to be a potent nucleic-acid chaperone that assists RT during the conversion of the genomic RNA into proviral DNA. Here we briefly review the interplay of NC with viral nucleic-acids, which enables rapid and faithful folding and hybridization of complementary sequences, and with active RT thus providing assistance to the synthesis of the complete proviral DNA. Because of its multiple roles in retrovirus replication, NC could be viewed as a two-faced Janus-chaperone acting on viral nucleic-acids and enzymes.

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