Viruses

General terms of Viruses

• Virus: genetic element that cannot replicate independently of a living
(host) cell . Particles of nucleic acid, protein, and sometimes lipids

• Virology: the study of viruses

• Virus particle: extracellular form of a virus; allows virus to exist outside

host and facilitates transmission from one host cell to another

• Virion: the infectious virus particle; the nucleic acid genome surrounded
by a protein coat and, in some cases, other layers of material

• Viral Genomes

– Either DNA or RNA genomes

– Some circular, but most linear

 Novel properties of viruses
• Are obligate intracellular parasites of bacteria, protozoa, fungi, algae, plants and
animals
• Are not cells; structure is very compact
• Do not independently fulfil the characteristics of life
• Are inactive macromolecules outside of the host cell and active only inside host
cell
• Are geometric; can form crystal like masses
• Basic structure consists of protein shell (capsid) surrounding nucleic acid core
• Nucleic acid can be either DNA or RNA but not both
• Nucleic acid can be double stranded DNA, single stranded DNA, single stranded
RNA or double stranded RNA
• Molecules on virus surface impart high specificity for attachment to host cell
• Multiply by taking control of host cell's genetic material and regulating the
synthesis and assembly of new viruses
• Lack enzymes for most metabolic processes
• Lack machinery for synthesizing proteins
Size of viruses

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 Host range
• Host range refers to the spectrum of host cells in which a virus can multiply.
• Host range is determined by the specific attachment on the host cell's surface
and the availability of host cellular factors. Viruses can be classified on the
basis of the hosts they infect
• Bacterial viruses (bacteriophages)
• Animal viruses
• Plant viruses

Viral Size
• Viral size is ascertained by electron microscopy
• Viruses range from 20 to 1000 nm in length
Viral Structure
A virion is a complete, fully developed viral particle composed of
nucleic acid surrounded by a coat.
 Nucleic acid
• Viruses contain either DNA or RNA. Never both, and the nucleic acid may
be single or double stranded, linear or circular, or divided into several
separate molecules.
• The proportion of nucleic acid in relation to protein in viruses ranges from
about 1% to about 50%

Capsid and Envelope

• The protein coat surrounding the nucleic acid of a virus is called the
capsid.
• The capsid is composed of subunits, capsomeres, which can be a single
type of protein or several types
• 2 types: helical and iscosahedral
• The capsid of some viruses is enclosed by an envelope consisting of lipids,
proteins and carbohydrates
• Some envelopes are covered with carbohydrate-protein complexes called
spikes.
Capsid morphology:
• Helical: Ribbon-like protein forms a spiral around the
nucleic acid. May be rigid or flexible.
– Tobacco mosaic virus
– Ebola virus
• Polyhedral: Many-sides. Most common shape is
icosahedron, with 20 triangular faces and 12 corners.
– Poliovirus
– Herpesvirus
• Complex viruses: Unusual shapes
– Bacteriophages have tail fibers, sheath, and a plate
attached to capsid.
– Poxviruses have several coats around the nucleic acid.
helical
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icosahedral
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icosahedral
• 20-sided with 12 corners
• Vary in the number of
capsomeres
• Each capsomere may be
made of 1 or several
proteins
• Some are enveloped

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complex
Viruses are classified by the following
characteristics:
– Type of genetic material
– Capsid shape
– Number of capsomeres
– Size of capsid
– Presence or absence of envelope
– Host infected
– Type of disease produced
– Target cell
– Immunological properties
 Taxonomy of Viruses

• Classification of viruses is based on type of nucleic acid,

strategy for replication, and morphology.
• Virus family names end in -viridae; genus names end in -
virus.
• A viral species is a group of viruses sharing the same
genetic information and ecological niche.
 Viral Reproduction

• Requires host cell

• Bacteriophages (prokaryotes)

• Animal viruses (eukaryotes)

BACTERIOPHAGES
 Bacteriophages Reproduction Cycle
• Lytic Cycle
– Mass production of phage
– Ends in host cell death (lysis)

• Lysongenic cycle
– Virus reproduces through binary fission along with host
cell
– Can spontaneously convert to lytic cycle
 Bacteriophage Reproduction cycle

• Adsorption/ Attachment
– Random encounter
– Attachment – Receptor connection (lock & key)
– The tail attaches to the cell surface and the tail pins
protrude from the base of the tail.

• Penetration
– Break down of cell wall (phage lysozyme)
– Injection of DNA into cytoplasm
– The protein coat, which forms the phage head and the tail
structures of the virus remain outside the cell.
Penetration

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 Bacteriophage Reproduction cycle (Lytic)
• Transcription
– Viral DNA takes over. Bacterial DNA polymerase
helps in transcription of new phage genes.
– Host metabolic machinery synthesizes new viral
parts (heads, tails and fibers). They include a
phage lysozyme which will lyse the bacterial cell,
releasing the mature virions.
• Assembly and Release
– After synthesis of both structural proteins and
nucleic acids, the phage components begin to
assemble into mature phages.
– Break down of host cell wall
– New virion escape to infect more cells
Burst time: Time from attachment to
release of new virions (20-40 minutes).

Burst size: Number of new phage

particles that emerge from a single cell
(50-200).
Bacteriophage assembly line

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Bacteriophage Lambda: Lysogenic Cycle
1. Attachment and Penetration: Virus tail binds to
specific receptors on the cell surface and injects
genetic material (DNA) into cell.
2. Circularization: Phage DNA circularizes and
enters either lytic or lysogenic cycle.
Lysogenic Cycle
3. Integration: Phage DNA integrates with
bacterial chromosome and becomes a
prophage. Prophage remains latent.
4. Excision: Prophage DNA is removed due to a
stimulus (e.g.: chemicals, UV radiation) and
initiates a lytic cycle.
Lysogenic versus Lytic Cycles of Bacteriophage
 Examples of Bacteriophage
• Bacteriophage T1, T4, T5,

• Coliphage lambda
 Isolation, Cultivation, and
Identification of Viruses
• Viruses must be grown in living cells.

• The most usual sources of bacteriophages is the host habitat.

• Bacterial viruses are easily isolated and cultivated in young,

actively growing cultures of bacteria in broth or on agar
plates.

• In liquid cultures, lysing of the bacteria will cause a cloudy

culture to become clear.

• In agar-plate culture, clear zones or plaques become visible to

the unaided eye.
 Quantification of Viruses
• Titer: number of infectious units per volume of fluid

• Plaque assay: analogous to the bacterial colony; one of the

most accurate ways to measure virus infectivity.

• Plaques are clear zones that develop on lawns of host

cells.

• Each plaque results from infection by a single virus particle

Quantification of Bacterial Virus by Plaque Assay

Figure 10.6
Quantification of Bacterial Virus by Plaque Assay

Figure 10.6
 Quantification of Viruses

• The number of plaque-forming units is

almost always lower than direct counts by
microscopy
– Inactive virions

– Conditions not appropriate for infectivity

 Introduction to Animal Virus
• Smallpox vaccine found out by Edward
Jenner.

• Variola (smallpox) virus introduced into

subject to provide protection against natural
smallpox infection.
• Smallpox vaccine is made from virus grown
on skin of healthy calves or sheep.

• Tobacco mosaic virus which causes tobacco

mosaic disease on tobacco leaves.
 Structure and Composition
• Composed of central core of nucleic acid surrounded by
capsid which is made up of capsomeres.
• Nucleocapsid is sometimes covered with an envelope.
• Most plant and animal viruses exhibit a characteristic
symmetry.
– Icosahedral in spherical viruses: polioviruses (poliomyelitis)
and adenoviruses (respiratory infections)
– Helical in rod shaped viruses: tobacco mosaic virus,
measles, mumps, influenza and rabies viruses.
– Complex in miscellaneous group: pox virus
 Structure and Composition
• Nucleic acids:
• Other chemical components:
– Protein: Capsid is made up of protein. Internal proteins
may also be present in some viruses (adenovirus).
– Lipid: found in viral envelope. They may be in the form of
phospholipids, glycolipids, neutral fats, fatty acids,
cholesterol etc.
– Carbohydrate: nucleic acid itself contains ribose or
deoxyribose. Some enveloped animal viruses have spikes
made up of glycoprotein on envelope (influenza virus)
 Host range
• Spectrum of cells a virus can infect
– cell has to have a specific structure (receptor) on its
surface for viral attachment
– cell has to contain all of the enzymes and materials
needed to produce new virions
• May be one species or many
– HIV (only humans) vs rabies (many animals)
• May be one tissue or many within a host
– Hepatitis (liver) vs polio (intestinal & nerve cells)

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 Animal virus replication
• Virus particles outside the host cell have no
independent metabolic activity.
• They are thus incapable of reproduction by
processes characteristic to other
microorganisms.
• Multiplication takes place by replication.
• Reproduction of virus takes place in the
cytoplasm, the nucleus or both.
 Life Cycle of Animal Viruses
1. Attachment or adsorption: Virus binds to
specific receptors (proteins or glycoproteins) on
the cell surface.
2. Penetration: Virus enters cell through one of
the following processes:
– Direct fusion with cell membrane
– Endocytosis through a clathrin coated pit
3. Uncoating: Separation of viral nucleic acid from
protein capsid. Lysosomal, cytoplasmic, or viral
enzymes may be involved.
Attachment, Penetration, and
Uncoating of Herpes Virus
Life Cycle -Animal Viruses (Continued)
4. Synthetic Phase: Involves several processes:
• Synthesis of viral proteins in cytoplasm
• Replication of viral genome:
– DNA viruses typically replicate in nucleus
– RNA viruses replicate in cytoplasm
• Assembly of progeny virus particles
The synthetic stage can be divided in two periods:
• Early period: Synthesis of proteins required for
replication of viral genetic material.
• Late period: Nucleic acid replication and synthesis
of capsid and envelope proteins
Life Cycle-Animal Viruses (Continued)
5. Release of progeny virions: There are two main
mechanisms of release:
A. Lysis of cells: Naked viruses and pox viruses leave cell
by rupturing the cell membrane.
Usually results in death of the host cell.
Example: Poliovirus

B. Budding: Enveloped viruses incorporate viral

proteins in specific areas of a membrane and bud
through the membrane.
Envelope contains host lipids and carbohydrates.
Host cell does not necessarily die.
Example: Human Immunodeficiency Virus
adsorption

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penetration

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Release by budding

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 Differences between phage and
animal virus replication

1. Animal virus replication is more complex

than phage replication because host cells are
more complex.
2. Animal viruses cannot inject their DNA.
3. Lysogeny for phage, latency for animal
viruses

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 Bacteriophages and animal viral multiplication
compared

Bacteriophages
Animal
• Tail fibers attach to cell wall •Attachment sites are plasma membrane
proteins proteins and glycoproteins.
• Viral DNA injected into host
cell •Capsid enters by endocytosis or fusion
• Not required •Enzymatic removal of capsid proteins
• In cytoplasm
•In nucleus DNA viruses or cytoplasm RNA
viruses

• Lysogeny •Latency; slow viral infections

•Enveloped viruses bud out; noneveloped

• Host cell lysed viruses rupture plasma membrane.
 Isolation and Identification of Virus
• Proper collection and care of specimen until
susceptible animals, tissue cultures or
embryonated eggs are inoculated.

• If virus is present, characteristic antibodies,

complement-fixing or neutralizing viral
antibodies may be produced.
 Cultivation of Animal Virus
• Embryonated chicken eggs: the chick embryo technique has
been used in the production of vaccines against smallpox,
yellow fever, influenza and in immunologic tests and othe
studeies whenever large amounts of virus are required.

• Tissue cultures: it is a method for the propagation of viruses

because it is convenient, relatively economic. Cell lines can be
obtained from mouse, hamster, chicken or monkey tissue etc.

• Animals: living animals such as mice, guinea pigs and rabbits

are used. Animal inoculation is a good diagnostic tool since
they can show disease symptoms and tissue sections can be
used for microscopic examinations.
 Diagnosis of viral diseases
• More difficult than other agents
• Consider overall clinical picture
• Take appropriate sample
– Infect cell culture- look for characteristic
cytopathic effects
– Screen for parts of the virus
– Screen for immune response to virus (antibodies)

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Chickenpox (Varicella zoster virus)
• Chickenpox is one of the classic
childhood diseases, and one of
the most contagious. The
affected child or adult may
develop hundreds of itchy, fluid-
filled blisters that burst and form
crusts. Chickenpox is caused by a
virus.
 Latent Viral Infections
• A latent viral infection is one in which the virus remains in the
host cell for long periods without producing an infection.
• Examples are cold sores and shingles.

Persistent Viral Infections

• Persistent viral infections are disease processes that occur over a
long period and are generally fatal.
• Persistent viral infections are caused by conventional viruses;
viruses accumulate over a long period.