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Virus Example
Parvoviruses
X174, M13,fd phase
Herpes viruses, cytomegalovirus, Epstein
Barr virus, adenovirus, T-coliphages,
Double- Stranded
Rhabdoviruses(rabies),
paramyxovirus(Mumps measles)
2. Protein Capsid
Viral genomes are surrounded by protein shells known as capsids. A capsid is almost always
made up of repeating structural subunits that are arranged in one of two symmetrical structures,
a helix or an icosahedron. In the simplest case, these "subunits" consist of a single polypeptide.
In many cases, however, these structural subunits (also called protomers) are made up of
several polypeptides. Both helical and icosahedral structures are described in more detail below.
Capsid is protein coat and Capsomeres are subunits of the capsid. Protomeres are capsomere
subunits.
Helical Capsids: The first and best studied example is the plant tobacco mosaic virus
(TMV), which contains a SS RNA genome and a protein coat made up of a single, 17.5
kd protein. This protein is arranged in a helix around the viral RNA, with 3 nt of RNA
fitting into a groove in each subunit. Helical capsids can also be more complex, and
involve more than one protein subunit.
A helix can be defined by two parameters, its amplitude (diameter) and pitch, where pitch
is defined as the distance covered by each turn of the helix. P = m x p, where m is the
number of subunits per turn and p is the axial rise per subunit. For TMV, m = 16.3 and p=
0.14 nm, so P=2.28 nm. This structure is very stable, and can be dissociated and reassociated readily by changing ionic strength, pH, temperature, etc. The interactions that
hold these molecules together are non-covalent, and involve H-bonds, salt bridges,
hydrophobic interactions, and van-der Waals forces. Several families of animal virus
contain helical nucleocapsids, including the Orthomyxoviridae (influenza),
the Paramyxoviridae (bovine respiratory syncytial virus), and the Rhabdoviridae (rabies).
Icosahedral Capsids: In these structures, the subunits are arranged in the form of a
hollow, quasi spherical structure, with the genome within. An icosahedron is defined as
being made up of 20 equilateral triangular faces arranged around the surface of a
sphere. They display 2-3-5 fold symmetry as follows:
- An axis of 2 fold rotational symmetry through the center of each edge.
- An axis of 3 fold rotational symmetry through the center of each face.
- An axis of 5 fold rotational symmetry through the center of each corner.
These corners are also called Vertices, and each icosahedron has 12.
Since proteins are not equilateral triangles, each face of an icosahedron contains more than one
protein subunit. The simplest icosahedron is made by using 3 identical subunits to form each
face, so the minimum of subunits is 60 (20 x 3).
Many viruses have too large a genome to be packaged inside an icosahedron made up of only 60
polypeptides (or even 60 subunits), so many are more complicated. In these cases, each of the 20
triangular faces is divided into smaller triangles; and each of these smaller triangles is defined by
3 subunits. However, the total number of subunits is always a multiple of 60. The total number of
subunits can be defined as 60 X N, where N is sometimes called the Triangulation Number, or
T. Values for T of 1,3,4,7,9, 12 and more are permitted.These are usually protein subunits
clustered around an axis of symmetry, and have been called "morphological units"
or capsomers.
3.Viral Envelope
In some animal viruses, the nucleocapsid is surrounded by a membrane, also called
an envelope. This envelope is made up of a lipid bilayer, and is comprised of host-cell lipids. It
also contains virally encoded proteins, often glycoproteins which are trans-membrane proteins.
These viral proteins serve many purposes, such as binding to receptors on the host cell, playing a
role in membrane fusion and cell entry, etc. They can also form channels in the viral membrane.
Many enveloped viruses also contain matrix proteins, which are internal proteins that link the
nucleocapsid to the envelope. They are very abundant (ie, many copies per virion), and are
usually not glycosylated. Some virions also contain other, non-structural proteins that are used
in the viral life cycle. Examples of this are replicases, transcription factors, etc. These nonstructural proteins are present in low amounts in the virion.
Enveloped viruses are formed by budding through cellular membranes, usually the plasma
membrane but sometimes an internal membrane such as the ER, golgi, or nucleus. In these cases,
the assembly of viral components (genome, capsid, matrix) occurs on the inside face of the
membrane, the envelope glycoproteins cluster in that region of the membrane, and the virus buds
out. This ability to bud allows the virus to exit the host cell without lysing, or killing the host. In
contrast, non-enveloped viruses, and some enveloped viruses, kill the host cell in order to escape.
Nature of genome
Example
Single stranded DNA
X174, fd
Double stranded DNA
phages , -phages
Single stranded RNA genome
Of plus sense
Doublestranded RNA
TMS2
6
Cell culture is a lot cheaper and easier to work with (contamination can be a problem however).
Primary cell lines have a short lifespan in culture a few generations before reaching
senescence. Diploid cell lines are derived from embryos and can grow for up to 100 population
doublings before senescence. Continuous cell lines are derived from transformed cells and grow
indefinitely in culture (Fig.).
Hela cells 1st continuous cell line, derived from Helen Lane (fictional name - actually named
Henrietta Lacks), a cervical cancer patient who died in 1951. This is the oldest continuous cell
line and was first used to culture and identify polio virus.
Viral Multiplication
Viruses do not contain enzymes for energy production or protein synthesis. For a virus to
multiply, it must invade a host cell and direct the hosts metabolic machinery to produce viral
enzymes, viral proteins, and copies of its nucleic acid, using the host cell's ATP to power the
reactions. Viral particles disappear upon penetration, none are seen during biosynthesis and
assembly, and eventually all cells die so no new virions can be produced. The eclipse period is
the period when all viral particles are present but before they are assembled.
It is a cycle in which the phage DNA recombines with the bacterial chromosome. The
incorporated viral DNA is now a prophage. The prophage genes are regulated by a repressor
coded for by the prophage, the prophage is replicated each time the host DNA is replicated.
Exposure to mutagens can lead to excision of the prophage and initiation of the lytic cycle.
Table: Bacteriophage and viral multiplication compared
Oncogenic viruses are capable of producing tumors in animals. Oncogenic viruses integrate into
host cell DNA and may cause transformation of host cells.Transformed cells lose contact
inhibition, contain virus-specific antigens (tumor-specific transplantation antigen (TSTA)on the
cell surface or T antigen in the nucleus), exhibit chromosomal abnormalities, and can produce
tumors when injected into susceptible animals.
Tumor Types:
Benign - tumors that generally dont spread, can be removed, and arent life threatening
Malignant invasive, generally aggressive tumors that are life threatening
Characteristics of Benign and Malignant Tumors
Organization (differentiation and anaplasia)
Differentiation: the extent to which tumor cells resemble the cell of origin in both
appearance and function
Anaplasia: undifferentiated, cells appear almost embryonic
Pleomorphism: variation in shape and size
Dysplasia: disorganized but non-neoplastic
The relationship between anaplasia and growth rate and specialized function is inverse the more
anaplastic and the faster growing cells are the less likely to have specialized function. Benign
tumors tend to be more differentiated, exhibit dysplasia, and the cells aren't usually pleomorphic.
Malignant tumors tend to exhibit anaplasia and pleomorphism.
Rate of growth
Benign tumors generally grow slowly. Malignant tumors exhibit a correlation between rate of
growth and degree of differentiation. May have periods of slow growth followed by rapid
growth, may spontaneously regress to due central necrosis and inability to provide nutrition
Local invasion
Benign tumors are usually demarcated and often encapsulated. Malignant tumors are invasive;
penetrate surrounding tissue
Metastasis
Development of secondary tumors distant from the site of the original tumor, characteristic of
malignant tumors
Fig. Process of prions toward to normal to abnormal form and result is cell death
Diseases:
Scrapie (sheep)
Transmissible mink encephalopathy
Bovine spongiform encephalitis (mad cow disease)
Kuru New Guinea, contracted by eating infected brain tissue (cannabilism was a
mourning rite among members of this particular tribe between about 1920 and 1950)
Creutzfeldt-Jakob disease (CJD) is very similar and has a heritable form and may be
passed by contact (neurosurgery, corneal transplants, pituitary-derived GH preparations)
Gerstmann-Straussler-Scheinker syndrome (GSS) similar, inherited
Plant Viruses and Viroids
Examples:
Viroids are naked (lacking a protein coat) pieces of RNA that can cause some plant diseases.
They are internally base paired, so they assume a folded conformation that protects them from
enzymatic degradation.
Viroids don't code for proteins and research indicates that they have similarities to introns, which
suggests researchers may discover animal viroids in the future.
Example: Potato spindle tuber viroids
Diseases With Possible Viral/Prion Etiology
of the
Fig. Anatomy
AIDS Virus
HIV
genome:
HIV-1 is
a retrovirus. It
contains two copies of plus single stranded RNA genome. Most retroviruses that are capable of
replication contain only three genes namely gag, pol and env. HIV-1 contains nine different
genes in its 9kb RNA genome (gag, pol, env, vif, vpu vpr, tat, rev and nef). Flanking these genes
are the Long Terminal Repeats (LTR), Which contain regulatory elements involved in gene
expression. The major difference between the genome of HIV-1 and HIV-2 is that HIV-2 lacks
the vpu gene and has a vpx gene not contained in the HIV-1.
Gene
gag
pol
env
vif
vpu
vpr
tat
rev
nef
2.
Budding: This is the final stage of the virus life cycle. In this stage, the virus pushes
itself out of the host cell, taking with it part of the membrane of the cell. This outer part
covers the virus and contains all of the structures necessary to bind to a new CD4 cell and
receptors and begin the process again.
These steps of the life-cycle of HIV are important to know because the medications used to
control HIV infection act to interrupt this replication cycle.
HIV-1 infects T-cells that carry the CD4 antigen on their surface; in addition certain HIV strains
will infect monocytes and other cells that have CD4 on their surface. Binding to CD$ on the cell
surface and viral entry is also mediated by Chemokine co-receptors CXCR4 or fusin and CCR5
present on T-cell and monocytes, respectively.
After the virus has entered the cell, the RNA genome of the virus is reverse transcribed and
translate into proteins, which along with a complete new copy of the RNA genome are used to
form new viral particles.
Infection of target cell:
Activation of provirus:
Transcription factors stimulate transcription of proviral DNA into genomic ssRNA and
after processing several mRNAs.
Viral RNA is exported to the cytoplasm.
Host-cell ribosomes catalyze the synthesis of viral precursor proteins.
Viral proteases cleave precursors into which gp41 and gp120 are inserted.
The membrane buds out, forming the viral envelope.
Released viral particles complete maturation; incorporated precursor protein are cleaved
by the viral protease present in viral particles.
Therapeutic agents:
At present there is no complete cure for AIDS. Primary treatment is directed at reducing the viral
load and disease symptoms, and at treating opportunistic infection and malignancies. The
antivirals currently approved for use in HIV disease are of two types.
Reverse transcriptase inhibitors are nucleoside analogous that inhibit the enzyme reverse
transciptase as it. Synthesizes DNA from RNA. Examples include AZT or zidovudine (Retrovir),
didanosine(Videx), zalcitabine(HIVID), stavudine, lamivudine, delavirdine and nevirapine
( viramune).
Protease inhibitors work by blocking the activity of the HIV protease and thus interfere with
virion assembly. Examples include indinavir (Crixivan), ritonavir, nelfinavir( viracept), and
saquinavir (Invirase). However, the most successful treatment approach is to use drug
combinations. An effective combination is a cocktail of AZT, lamirudine, and a protease inhibitor
such as ritonavir.
Stages of HIV infection:
Antiretroviral therapy (ART) prevents the HIV virus from multiplying and from destroying your
immune system.
a.Acute infection stage
Within 2-4 weeks after HIV infection, many people develop flu-like symptoms can include fever,
swollen glands, sore throat, rash, muscle and joint aches and pains, fatigue, and headache. This is
called acute retroviral syndrome (ARS) or primary HIV infection, and its the bodys natural
response to the HIV infection.
b.Clinical latency stage
After the acute stage of HIV infection, the disease moves into a stage called the clinical latency
stage. Latency means a period where a virus is living or developing in a person without
producing symptoms. During the clinical latency stage, people who are infected with HIV
experience no HIV-related symptoms, or only mild ones. (This stage is sometimes
called asymptomatic HIV infection or chronic HIV infection.)
During the clinical latency stage, the HIV virus continues to reproduce at very low levels,
although it is still active. If you take ART, you may live with clinical latency for several decades
because treatment helps keep the virus in check.
c.AIDS
This is the stage of HIV infection that occurs when your immune system is badly damaged and
you become vulnerable to infections and infection-related cancers called opportunistic infections.
When the number of your CD4 cells falls below 200 cells per cubic millimeter of blood (200
cells/mm3), you are considered to have progressed to AIDS. (In someone with a healthy immune
system, CD4 counts are between 500 and 1,600 cells/mm3.) As HIV disease progresses in your
body, you may notice physical changes. Some changes may occur as side-effects of medical
treatment for HIV. Others may occur as a result of the impact that HIV (or AIDS) has on your
body. The best treatment for HIV infection is Highly active antiretroviral therapy (HAART).