Nucleic acid viral

Short replication cycles that may be completed within a few hours, a large amount of viral progeny from one infected host-cell, as well as the general inaccuracy of viral nucleic acid polymerases result in an evolution occurring in fast motion, allowing rapid adaptation of viruses to selective pressures (see chapter by Boucher and Nijhius, this volume). Generalizing, it can be stated that any effective antiviral therapy will lead to the occurrence of resistance mutations. A well studied example... 

Virus maturation and assembly at the cell membrane or the nuclear membrane has long been seen as a potential target for antiviral compounds. For the virus to mature and be released in a conformation that will insure stability and survival of the viral genome in the exttacellular enviromnent, the protein subunits of the capsid or nucle-ocapsids have to be transported to the assembly point where they will form the final particles around the viral nucleic acid. If this process does not occur in an orderly and programmed manner, the capsid subunits will not form the required multimers and the viral components will become targets for the cellular disposal mechanisms. 

Peptoids have also shown great utility in their ability to complex with and deliver nucleic acids to cells, a critical step toward the development of antisense drugs, DNA vaccines, or gene-based therapeutics. Most non-viral nucleic acid delivery systems are based on cationic molecules that can form complexes with the polyan-... 

Small quantities of additional purines and pyrimidines occur in DNA and RNAs. Examples include 5-methyl-cytosine of bacterial and human DNA, 5-hydroxy-methylcytosine of bacterial and viral nucleic acids, and mono- and di-N-methylated adenine and guanine of... 

Helical symmetry replication of viral nucleic acid... 

A helical virus partially disrupted to show the helical coil of viral nucleic acid embedded in the capsomeres... 

In general terms, four main stages can be recognized in the multiplication of human viruses, (i) attachment (ii) penetration and uncoating (iii) production of viral proteins and replication of viral nucleic acid, (iv) assembly and release of progeny viruses. 

The non-enveloped human viruses all have icosahedral capsids. The structural proteins undergo a self-assembly process to form capsids into which the viral nucleic acid is packaged. Most non-enveloped viruses accumulate within the cytoplasm or nucleus and are only released when the cell lyses. 

Wilber, J. C., and Urdea, M. S. (1995). Chapter 6. In Molecular methods for virus detection. Quantification of viral nucleic acids using branched DNA signal amplification. (San Diego Academic Press). 

Enzymes in viruses We have stated that virus particles do not carry out metabolic processes. Outside of a host cell, a virus particle is metabolically inert. However, some viruses do contain enzymes which play roles in the infectious process. For instance, many viruses contain their own nucleic acid polymerases which transcribe the viral nucleic acid into messenger RNA once the infection process has begun. The retroviruses are RNA viruses which replicate inside the cell as DNA intermediates. These viruses possess an enzyme, an RNA-dependent DNA popo called reverse transcriptase, which transcribes the information in the incoming RNA into a DNA intermediate. It should be noted that reverse transcriptase is unique to the retroviruses and is not found in any other viruses or in cells. 

As we have noted, the outcome of a virus infection is the synthesis of viral nucleic acid and viral protein coats. In effect, the virus takes over the biosynthetic machinery of the host and uses it for its own synthesis. A few enzymes needed for virus replication may be present in the virus particle and may be introduced into the cell during the infection process, but the host supplies everything else energy-generating system, ribosomes, amino-acid activating enzymes, transfer RNA (with a few exceptions), and all soluble factors. The virus genome codes for all new proteins. Such proteins would include the coat protein subunits (of which there are generally more than one kind) plus any new virus-specific enzymes. 

Virus restriction and modification by the host We have already seen that one form of host resistance to virus arises when there is no receptor site on the cell surface to which the virus can attach. Another and more specific kind of host resistance involves destruction of the viral nucleic acid after it has been injected. This destruction is brought about by host enzymes that cleave the viral DNA at one or several places, thus preventing its replication. This phenomenon is called restriction, and is part of a general host mechanism to prevent the invasion of foreign nucleic acid. 

Subsequently, similar experiments were done with viral nucleic acids. The pure viral nucleic acid, when added to cells, led to the synthesis of complete virus particles the protein coat was not required. This process is called transfection. More recently, DNA has been used in cell-free extracts to program the synthesis of RNA that functions as the template for the synthesis of proteins characteristic of the DNA... 

Penetration. After fusion of viral and host membranes, or uptake into a phagosome, the virus particle is carried into the cytoplasm across the plasma membrane. This penetration process is an active one that requires expenditure of energy by the cell. At this stage the envelope and the capsid are shed, and the viral nucleic acids are released. The uncoating of virus accounts for the drop in infectious virus assayed, because the uncoated virus cannot withstand the assay conditions. 

Translation of Viral mRNA. Once viral mRNA has been formed, translation occurs in the host cytoplasm, using host ribosome to synthesize viral proteins. Viral mRNA, which is usually monocistronic (i.e., has a single coding region) can displace host mRNA from ribosome so that viral products are synthesized preferentially. In the early phase, the proteins produced (enzymes, regulatory molecules) are those that will allow subsequent replication of viral nucleic acids in the later phase, the proteins necessary for the formation of capsid are produced. 

Replication of Viral Nucleic Acid. In addition to producing molecules for the formation of new capsids, the virus must replicate its nucleic acid to provide genetic material for packaging into the capsids. The way in which this is done might vary. In positive-sense, single-strand RNA viruses, a polymerase translated from viral mRNA produces negative-sense RNA from the positive-sense template which is then repeatedly transcribed into more positive strands. 

Structure and function of virus proteins and of viral nucleic acid. In The Proteins Composition, Structure, Function", Vol. 3, 99—151. Ed. II. Neurath, Academic Press, 2i,d Ed. 1965. 

Both the ELISA and Western blot suffer from the problem that antibodies may not appear in an aposed individual s blood until months after the initial exposure. Methods for using PCR to screen blood samples for HIV are being developed, PCR amplification of the HIV provirai DNA 7-ovides the ability to detect HIV at earlier stages of infection, because the viral nucleic acid is r resent immediately upon exposure. It is used to detect HIV infection in newborns whose mothers are HIV positive. 

The use of antibiotics for the control of plant virus diseases( ) is of interest. Several antibiotics have been tested for inhibition of replication of viral nucleic acid and/or protein synthesis within the host cell. Chloramphenicol, cycloheximide, actinomycin D and others are the most used antibiotics and the disease caused by tobacco mosaic... 

Pharmacology Inhibits the replication of influenza A virus isolates from each of the subtypes. Amantadine s antiviral activity is not completely understood. Its mode of action appears to be the prevention of the release of infectious viral nucleic acid P.1044... 

Figure 9.17. Overview of the manufacture of Ceprotin. As the active ingredient is derived directly from pooled human plasma, particular emphasis is placed upon ensuring that the finished product is pathogen-free. Precautions entail the incorporation of two independent viral inactivation steps and high-resolution chromatographic purification. Additionally, extensive screening of plasma pool source material for blood-borne pathogens is undertaken. Viral screening is undertaken using a combination of immunoassay and PCR analysis for the presence of viral nucleic acid...

Uncoating of the viral nucleic acid through shedding of the protein coat... 

Viral DNA polymerase is an important catalyst for the synthesis of viral nucleic acids. DNA polymerase inhibitors have already been encountered as antitumor agents. Ara-A (9.5, vidarabine) is a DNA polymerase inhibitor that has demonstrated activity against herpes simplex virus type I (HSV-1) infections, responsible for cold sores on... 

Viral replication consists of several steps (Figure 49-1) (1) attachment of the vims to receptors on the host cell surface (2) entry of the virus through the host cell membrane (3) uncoating of viral nucleic acid (4) synthesis of early regulatory proteins, eg, nucleic acid polymerases (5) synthesis of new viral RNA or DNA (6) synthesis of late, structural proteins (7) assembly (maturation) of viral particles and (8) release from the cell. Antiviral agents can potentially target any of these steps. 

Using the techniques described in Sections 1.1. and 1.2., it is possible to label both nucleic acids sequences and proteins on the same section. This has been used to colocalize viral nucleic acids and proteins (Fig. 5) (6,7). 

The majority of antiviral drugs which are under clinical development today generally interrupt viral nucleic acid synthesis. These compounds often do not affect host cell metabolism and possess considerable selectivity against virus-induced enzymes. This article discusses agents exhibiting significant antiviral activity against viral infections in animal model systems. 

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