Chapter 13: Viruses, Viroids,

and Prions

Ghaleb Adwan
 General Characteristics of Viruses

1. They are acellular, that is, they contain no cytoplasm or cellular

organelles.
2. Viruses contain:
- a single type of nucleic acid (DNA or RNA)
- a protein coat
- sometimes enclosed by an envelope encoded by the host cell
genome composed of:
- lipids, proteins, carbohydrates
3. They are totally dependent on a host cell for replication.
(obligatory intracellular parasites), multiply inside living cells
4. Most viruses are able to infect specific types of cells of only one
host species.
5. small (filterable)
 Host Range
• Host range of a virus: the spectrum of host cells the
virus can infect.
• There are viruses that infect invertebrates,
vertebrates, plants, protists, fungi, and bacteria.
• However, most viruses are able to infect specific
types of cells of only one host species
• In rare cases, viruses cross the host-species barrier,
thus expanding their host range.
• particular host range of a virus is determined by the
virus’s requirements for its specific attachment to the
host cell and the availability within the potential host of
cellular factors required for viral multiplication
 Viral Size
• Different viruses vary considerably in size.
• most are quite a bit smaller than bacteria, some of the
larger viruses (such as the vaccinia virus) are about
the same size as some very small bacteria (such as
the mycoplasmas, rickettsias, and chlamydias).
• Viruses range from 20 to 1000 nm in length.
Virus sizes. The sizes of several viruses (teal blue) and bacteria (brown) are
compared with a human red blood cell, shown below the microbes.
Dimensions are given in nanometers (nm) and are either diameters or length
by width.
 Viral Structure
• Virion: a complete, fully developed, infectious viral
particle composed of nucleic acid and surrounded by
a protein coat outside a host cell.
1. Nucleic Acid:
- either DNA or RNA, never both
-may be single- or double-stranded
- linear or circular
- or segmented
• total amount of nucleic acid varies from a few
thousand nucleotides (or pairs) to as many as 250,000
nucleotides
2. Capsid and Envelope:
• The protein coat surrounding the nucleic acid of a virus is called
the capsid.
• The capsid is composed of subunits called capsomere
• The arrangement of capsomeres is characteristic of a particular
type of virus
•

Morphology of a nonenveloped polyhedral virus. A diagram of a polyhedral

(icosahedral) virus.
• The capsid of some viruses is enclosed by an envelope
consisting of lipids, proteins, and carbohydrates.
• Some envelopes contain carbohydrate-protein complexes called
spikes or projections

Morphology of an enveloped helical virus. A diagram of an enveloped

helical virus.
• viruses attach to host cells by means of spikes
• Spikes can be used as a means of identification
• Viruses whose capsids aren’t covered by an envelope are known
as nonenveloped viruses
General Morphology
• Helical Viruses: long rods that may be rigid or flexible. The viral
nucleic acid is found within a hollow, cylindrical capsid that has a
helical structure
• viruses that cause rabies and Ebola are helical viruses
 Morphology of a helical virus. (a) A diagram of a portion of a helical virus. A row of
capsomeres has been removed to reveal the nucleic acid. (b) A micrograph of Ebola virus
2. Polyhedral viruses (e.g. adenovirus and poliovirus) are
many-sided. Usually the capsid is an icosahedron, with
20 equilateral triangle faces, 12 corners and 30 edges

Morphology of a nonenveloped polyhedral virus. (a) A diagram of a polyhedral (icosahedral)

virus. (b) A micrograph of the adenovirus Mastadenovirus.
3. Enveloped viruses: enveloped viruses are roughly spherical.
When helical or polyhedral viruses are enclosed by envelopes,
they are called enveloped helical or enveloped polyhedral
viruses.

Figure 3 Morphology of an enveloped helical virus

 4. Complex viruses: neither helical nor icosahedral
bacteriophages have a polyhedral capsid to which additional structures are attached
The capsid (head) is polyhedral and that the tail sheath is helical. The head
contains the nucleic acid.
Other structures as the tail fibers, baseplate, and pin
Another example of complex viruses are poxviruses,
which don’t contain clearly identifiable capsids but do
have several coats around the nucleic acid

Morphology of complex viruses

 Taxonomy of Viruses
*The International Committee on Taxonomy of Viruses (ICTV)
lays down the rules for the nomenclature and classification of
viruses, and it considers proposals for new taxonomic groups and
virus names.
1.The oldest classification of viruses is based on
symptomatology. Not scientifically acceptable because the same
virus may cause more than one disease, depending on the tissue
affected
2.Classification of viruses is based on:
- type of nucleic acid
- strategy for replication
- morphology
3. Then how their mRNA is produced.
4. Now taxonomy depends on DNA sequences
Baltimore classification of viruses
 Isolation and Cultivation of Viruses
Bacteriophages can be grown either in suspensions of bacteria in
liquid media or in bacterial cultures on solid media
solid media makes it possible to use the plaque method to detect
and count viruses.
• In the plaque method: bacteriophages are mixed with host bacteria
and melted nutrient agar, the virus-bacteria mixture solidifies into a
thin top layer in plate.
• After several viral multiplication cycles, the bacteria in the area
surrounding the original virus are destroyed; This produces a number
of clearings (plaques), visible against a lawn of bacterial growth on
the surface of the agar.
• Each plaque originates with a single viral particle; the
concentration of viruses is given as plaque-forming units (CFU).

Viral plaques formed by bacteriophages. Clear viral plaques of various sizes

have been formed by bacteriophage λ (lambda) on a lawn of E. coli.
 Growing Animal Viruses in the Laboratory
Animal Viruses can be grown in
1. Living Animals: Animal inoculation may be used as a diagnostic procedure
for identifying and isolating a virus from a clinical specimen. Some human
viruses can’t be grown in animals or can be grown but don’t cause disease.
2. Embryonated Eggs: convenient and inexpensive form of host for many
animal viruses. A hole is drilled in the shell of the embryonated egg, and a
viral suspension or suspected virus-containing tissue is injected into the
egg’s fluid.

Inoculation of an embryonated egg. The viruses will grow on the membrane at the inoculation site.
• 3. Cell Cultures
• Cell cultures have replaced embryonated eggs as the preferred type of
growth medium for many viruses
• Cell cultures are cells growing in culture media in the laboratory
• homogeneous collections of cells and can be propagated and handled much
like bacterial cultures, they are more convenient to work with than whole
animals or embryonated eggs.
• Viruses infecting such a monolayer sometimes cause the cells of the
monolayer to deteriorate as they multiply.
• This cell deterioration is called cytopathic effect (CPE).
• Primary cell lines: derived from tissue slices, tend to die out after only a few
generations.

Cell cultures. Transformed cells can be grown indefinitely in laboratory culture

• Diploid cell lines: developed from human embryos can be maintained
for about 100 generations and are widely used for culturing viruses that
require a human host (to culture rabies virus for a rabies vaccine called
human diploid culture vaccine)
• Continuous (immortal) cell lines: transformed (cancerous) cells that
can be maintained through an indefinite number of generations. HeLa
cell line, was isolated from the cancer of a woman (Henrietta Lacks).
There are still some viruses that have never been successfully cultivated
in cell culture.
Viral Identification
*Viruses can’t be seen without the use of an electron microscope.
*Serological methods, such as Western blotting, are the most commonly used
means of identification.
*Observation of cytopathic effects is a useful for identifying a virus
*modern molecular methods as use of restriction fragment length
polymorphisms (RFLPs) and the polymerase chain reaction (PCR)
 Multiplication of Bacteriophages
• The multiplication of viruses can be demonstrated with a one-step growth curve.
• Eclipse Phase: Virus has entered cell, but no progeny virions are created yet
• Maturation Phase: Viral progeny are produced and infectious virions are again present.
According to biosynthesis sequence
• Early Phase: Biosynthesis of prerequisite proteins, required before replication can take place,
such as replicase enzymes or enzymes that inhibit host-cell synthesis.
• Late Phase: Synthesis of structural capsid proteins, and actual replication of the genome.
 lytic cycle and lysogenic cycle
• Bacteriophages can multiply by two alternative mechanisms: the lytic cycle (ends
with the lysis and death of the host cell) or the lysogenic cycle (the host cell
remains alive).
• T-Even Bacteriophages: The Lytic Cycle
• T-even bacteriophages: complex, and nonenveloped, with a characteristic head-
and-tail structure.
• the phage has enough DNA for over 100 genes. The multiplication cycle of these
phages occurs in five distinct stages: attachment, penetration, biosynthesis,
maturation, and release.
1. Attachment (adsorption) phase: attachement sites on the phage's tail fibers
attach to receptor sites on the bacterial cell.
2. Penetration phase: bacteriophage’s tail releases a phage lysozyme, which
breaks down a portion of the bacterial cell wall, the tail sheath contracts to
force the tail core through the cell wall, and phage DNA enters the bacterial cell.
The capsid remains outside.
3. Biosynthesis: transcription of phage DNA produces mRNA coding for proteins
necessary for phage multiplication. The period during viral multiplication when
complete, infective virions aren’t yet present is called the eclipse period..
 4. Maturation: phage DNA and capsids are assembled into complete virions.
5. Release (lysis): phage lysozyme encoded by a phage gene, is synthesized
within the cell, breaks down the bacterial cell wall, and the multiplied phages
are released

Lytic cycle (T-even phage)

 Bacteriophage Lambda (λ): The Lysogenic
Cycle
• The lysogenic phages (temperate phages) may proceed through a
lytic cycle, but they are also capable of incorporating their DNA into
the host cell's DNA to begin a lysogenic cycle (the phage remains
latent (inactive)). The participating bacterial host cells are known as
lysogenic cells.
• Upon penetration of bacteriophage Lambda (λ) into an E. coli cell, the
linear phage DNA forms a circle. This circle can multiply and be
transcribed, leading to the production of new phage and to cell lysis
(the lytic cycle).
• Circluar phage DNA can recombine with and become part of the
circular bacterial DNA (the lysogenic cycle). The inserted phage DNA
is now called a prophage.
• However, a rare spontaneous event, or the action of UV light or
certain chemicals, can lead to the excision of the phage DNA, and to
initiation of the lytic cycle.
•
The lysogenic cycle of bacteriophage λ in E. coli.
 There are three important results of lysogeny
1. The lysogenic cells are immune to reinfection by the
same phage (isn’t immune to infection by other
phage types)
2. Phage conversion: the host cell may exhibit new
properties. The bacterium Corynebacterium
diphtheriae, which causes diphtheria, this organism
can produce toxin only when it carries a lysogenic
phage, because the prophage carries the gene coding
for the toxin . The toxin produced by Clostridium
botulinum, which causes botulism, is encoded by a
prophage gene.
3. Lysogeny makes specialized transduction possible.
Specialized transduction: When a prophage is excised from its host chromosome, it can take with it a bit of the
adjacent DNA from the bacterial chromosome
 Multiplication of Animal Viruses
• The multiplication of animal viruses follows the basic pattern of bacteriophage
multiplication but has several differences, summarized in Table .
The Biosynthesis of DNA and RNA
Viruses Compared
 Retroviridae
• One genus of retrovirus, Lentivirus, includes the
subspecies HIV-I and HIV-2, which cause AIDS.
• The name retrovirus is derived from the first letters of
reverse transcriptase (Retro = backward). These
viruses carry reverse transcriptase, which uses the
viral RNA as a template to produce complementary
double-stranded DNA.
• The viral DNA is then integrated into a host cell
chromosome as a provirus.
• In oncogenic retroviruses, the provirus can also
convert the host cell into a tumor cell.
Multiplication and inheritance processes of the Retroviridae A retrovirus may become a provirus that
replicates in a latent state, and it may produce new retroviruses .
 Viruses and Cancer
• The Transformation of Normal Cells into Tumor
Cells
• anything that can alter the genetic material of a
eukaryotic cell has the potential to make a normal cell
cancerous
• When activated, oncogenes transform normal cells into
cancerous cells.
• Viruses capable of producing tumors are called
oncogenic viruses or oncoviruses
• the cancer-inducing genes carried by viruses are
actually derived from animal cells.
• the cancer-causing src gene in avian sarcoma viruses is
derived from a normal part of chicken genes.
• Several DNA viruses and retroviruses are oncogenic.
• Transformed cells:
-viral DNA sequences present, integrated into
cellular DNA or as episome
-Greater growth potential in vitro
- lose contact inhibition
- exhibit chromosome abnormalities
- and can produce tumors when injected into
susceptible animals
-Virus-specified tumor-associated antigens
-cell surface behave as tumor-specific
transplantation antigens
-Altered cell morphology
-Altered cell metabolism and membrane changes
• Association of viruses with human cancer:
• A. DNA viruses
• 1. (Papovaviridae) Human papilloma viruses (HPV);
carcinoma of skin and ginitalia (Cervical carcinoma),
oropharyngeal carcinoma
• 2. (Herpesviridae) Epstein-Barr virus (EBV); Burkitt's
lymphoma ; Nasopharyngeal carcinoma; B-cell lymphoma
• 3. (Herpesviridae) Kaposi's sarcoma herpesvirus (HHV-8);
Kaposi's sarcoma
• 4. (Hepadnaviridae) Hepatitis B (HBV) primary liver cancer
(Hepatocellular carcinoma
• B. RNA viruses
• 1. (Retroviridae) Human T-cell lymphotropic
virus (HTLV-1); Adult cell leukemis/lymphoma
• 2. (Retroviridae) HIV; B-cell lymphoma,
invasive cervical carcinoma
• 3. (Flaviviridae) Hepatitis C (HCV); primary liver
cell cancer
 Prions

• Prions (proteinaceous infectious particle): infectious proteins

caused a neurological disease in sheep called scrapie. Several
Prion diseases included bovine spongiform
encephalopathy (BSE or mad cow disease). All are
neurological diseases called transmissible spongiform
encephalopathies because large vacuoles develop in the brain.
• The human diseases are kuru, Creutzfeldt-Jakob disease
(CJD), Gerstmann-Sträussler-Scheinker syndrome, and fatal
familial insomnia.
• all prion caused diseases
– have no effective treatment
– result in progressive degeneration of the brain and eventual death
• These diseases are caused by the conversion
of a normal host glycoprotein called PrPC (for
cellular prion protein) into an infectious form
called PrPSc (for scrapie protein). The gene for
PrPC is located on chromosome 20 in humans.
Recent evidence suggests that PrPC is involved
in preventing cell death.
 Viroids
• Viroids: short pieces of naked RNA, only 300 to 400
nucleotides long, with no protein coat. The nucleotides are
often internally paired, so the molecule has a closed,
folded, three-dimensional structure that presumably helps
protect it from destruction by cellular enzymes
• Some viroids, called virusoids (satellite RNA viauses),
are enclosed in a protein coat. Virusoids cause disease
only when the cell is infected by a virus (helper virus for
replication).
– human hepatitis D (Delta) virus is virusoid which require
human hepatitis B virus
• Viroids and virusoids are replicated continuously by host
RNA polymerase in the cell nucleus or chloroplasts
• The RNA doesn’t code for any proteins and
may cause disease by gene silencing
• Similarities between the base sequences of
viroids and introns, may viroids evolved from
introns
• cause plant diseases as potato spindle tuber
viroid