Virus Lecture

Copyright 2004 Pearson Education, Inc.
, publishing as Benjamin Cummings

Viruses, viroids and Prions
Copyright 2004 Pearson Education, Inc., publishing as Benjamin Cummings
Viruses
Obligatory intracellular parasites
Multiply inside living cells by using the synthesizing

machinery of the cell (have few or no enzymes)
Exist as non living entity outside living cell
Cause the synthesis of specialized structures that can transfer

the viral nucleic acid to other cells
Consists of
D! or "!
a protein coat
Host Range
Most viruses infect only specific types of cells in one host
#osts include
$nvertebrates
vertebrates
%lants
&ungi
'acteria (Bacteriophages, or phages)

Size of virus
(iruses range from )* to +*** nm in length
Can be seen only by electron microscope
most are smaller than bacteria
,ome are almost similar to bacteria
Eg-. vaccinia virus

Viruses
Viral Structure
Virion:
a complete. fully developed. infectious viral particle
composed of nucleic acid and surrounded by a protein coat

(Capsid)
/enome
D! or "!
single or double stranded
linear or circular
Capsid
protein coat
%rotects nucleic acid from environment
(ehicle of transmission from one to another host
Each capsid is composed of protein subunits called

capsomeres
single or several types of protein

Envelope
$n some viruses. the capsid is covered by an envelope
consists of lipid. protein and carbohydrates
0ayer of host cell plasma membrane for animal virus
$n some cases. envelop proteins determined by viral nucleic

acid
Spikes
Carbohydrate1protein complexes
pro2ect from the surface
3sed for attachment by some
3sed for identification

General Morphology
different morphological types on the basis of their capsid

architecture
1. Helical Viruses
long rods that may be rigid or flexible
4he viral nucleic acid is found within a

hollow. cylindrical capsid that has a
helical structure
Example5 "abies virus

Polyhedral Viruses
capsid of most polyhedral viruses is

in the shape of an icosahedron
a regular polyhedron with )*

triangular faces and +) corners
4he capsomeres of each face

form an e6uilateral triangle
Enveloped viruse
7hen helical or polyhedral viruses are

enclosed by envelopes. they are called
enveloped helical or enveloped
polyhedral viruses
roughly spherica0
enveloped helical virus is the influenza virus
enveloped polyhedral (icosahedral) virus is the herpes

simplex virus
Comple Viruses
8
(iruses that have complicated structures
8
Example5 bacteriophage
8
,ome have capsids to which additional
structures are attached
Classification of virus
(iruses are classified by differences in the nucleic !cid

1. Double-stranded D!: %ox virus
". Single-stranded D!: %arvovirus
#. Double-stranded $!: "otavirus
%. Single-stranded $!: #epatitis !
Viral !aonomy
&amily names end in 1viridae
/enus names end in 1virus
(iral species
! group of viruses sharing the same genetic information and

host
,ubspecies are designated by a number

(iruses are grouped into &amilies based on
+)nucleic acid type
))strategy for replication
9) morphology
Viral !aonomy
&amil': #erpesviridae
(enus: #erpesvirus
Species: #uman herpes virus
Subspecies: ##(+. ##( ). ##( 9

4he basic mechanism is similar for all virus
Entry and exit might vary
4o multiply virus needs to infect a host and use its

machinery
Most enzymes are obtained from the host
&rom a single virion several or even thousands of similar

viruses can develop in a single host cell
'acteriophages can multiply by two alternative mechanisms5

1. the l'tic c'cle: lysis and death of the host cell
". the l'sogenic c'cle: host cell remains alive
Multiplication of "acteriophages
#ytic Cycle $!%even
&acteriophage'
!ttachment
%enetration
'iosynthesis
Maturation
"elease
!ttachment %hage attaches by tail fibers to

host cell
%enetration %hage lysozyme opens cell wall.

tail sheath contracts to force tail
core and D! into cell
'iosynthesis %roduction of phage D!

and proteins
Maturation !ssembly of phage particles
"elease %hage lysozyme brea:s cell wall

Attachment:
Phage attaches
to host cell.
Penetration:
Phage penetrates
host cell and
injects its DNA.
3
Bacterial
cell wall
Bacterial
chromosome
Capsid
Sheath
Tail fiber
Base plate
Pin
Cell wall
Tail
Plasma membrane
Sheath contracted
Tail core
1.!ttachment ) !dsorption
&ollowed by chance collision
specific association;interaction of virion with specific receptors

on host cell surface (wea: bond forms)
'acterial mutant unable to synthesis specific receptors resistant

to infection by specific phage
susceptible cell contain ca- +*

<
receptor sites for a given virus
!bsorption highly specific for animal = bacterial virus but

nonspecific for plant virus (no specific receptors on host cell)
(irus might enter via phagocytosis or some other endocytic

process
". *enetration
entrance of viral D! into host cell cytoplasm
$n bacteriophages. more complicated than animal = plant viruses

steps in T even phages:
tail fibers contract
4ail core contacts envelope
0ysozyme li:e enzyme ma:es a hole
tail sheath contracts
tail core tube is forced into cell through

C7 = M (not obligatory for penetration)
D! passes through tube = into cytoplasm
protein coat remains outside

Bios'nthesis
$nside the cytoplasm. synthesis of viral nucleic acid and protein
#ost protein synthesis stopped
$nitially phage uses host>s nucleotides and enzymes for viral

nucleic acid synthesis
+aturation
assembly of capsid and nucleic acid
,pontaneous process
$elease
$n case of 41even phage lysozyme lyses cell wall
"eleased viruses infect susceptible cells in the vicinity
Multiplication continues
at!ration:
"iral components
are assembled into
#irions.
Tail fibers
Tail
DNA
Capsid
$elease: host cell
l%sis and new
#irions are
released
(ne%step Gro)th Curve
The period when complete virions cannot be detected in the cell
Is known as the eclipse period
Eclipse period $*n the first fe) minutes after infection'
!he viral nucleic acid &ecomes separated from its protein

coat
!he virion does not eist as an infectious entity
the infected cell &rea+s open

Maturation:
the ne)ly synthesized nucleic acid pac+aged inside protein

coat
the titre of active virions increase dramatically

Latent period:
time ta+en from penetration of ,- to &eginning of lysis $the
eclipse and maturation periods together'
replication of viral ,- . protein

/or !0 phage1
Eclipse1 20 min
#atent1 23 min $so latent is longer than eclipse'

Time for replication:
04%54 minutes for many &acteriophage
-nimal virus1 3%64 hours

Burst size:
!he yield of virion7host cell
varies )ith % virus

% type of host cell
% gro)th conditions
-verage yield1
for plant or animal cell1 several thousand to a million7Host Cell
/or &acteriophages1 84 9 2444

#ysogenic
Cycle
$#am&da
phage'
#ysogenic Cycle $#am&da
phage'
igure !".!2
*mportant Results of
#ysogeny
0ysogenic cells are immune to reinfection by the same phage (ot

immune to infection by other phage types)
%hage conversion1the host cell may exhibit new properties
e-g- Corynebacterium diphtheriae carries its toxin gene on a

lysogenic phage
Clostridium botulinum and Vibrio cholerae
,pecialized transduction is possible (pac:aging of bacterial D!

along with the viral D!)
Certain animal viruses can remain latent for long periods without multiplying or
%roducing disease
0ater the viral D! can be incorporated into the host chromosome
basic patterns of bacteriophage multiplication. with some

differences
!ttachment:
4o host cell membrane proteins or glycoproteins
o appendages li:e tail fibers
!ttachment sites are distributed all over the virus
(ary between viruses
!denovirus5 small fibers at the corners of the icosahedron
$nfluenza virus5 spi:es
&or host. receptor sites are inherited
(ary from person to person

Multiplication of -nimal viruses
Entr'
(iruses enter into the cell by endocytosis (pinocytosis)
%lasma membrane folds inward to form vesicles
Enveloped viruses can enter by fusion- Eg-. #$(

on1enveloped Enveloped
,ncoating
ucleic acid is separated from the capsid
Mediated by viral or host enzyme (varies between virus)
Capsid may be digested or released into host cell cytoplasm

Bios'nthesis
(ary between D! and "! virus

+aturation and release
3sually a spontaneous process
on enveloped viruses
migrate to the host>s cell membrane. rupture and release
Cell death
Enveloped viruses
released by budding
3sually cell survives

!ttachment 5 (iruses attaches to cell membrane

proteins (receptors are inherited)
%enetration5 'y endocytosis or fusion (enveloped

viruses)
3ncoating 'y viral or host enzymes
'iosynthesis %roduction of nucleic acid and proteins
Maturationucleic acid and capsid proteins assemble
"elease 'y budding (enveloped viruses) or

rupture
Multiplication of -nimal viruses
Enveloped Viruses
Examples5 $nfluenza virus and the herpes simplex virus
Generally, DNA-containing viruses replicate their DNA in the

nucleus of the host cell by using viral enzymes
Synthesize their capsid and other proteins in the cytoplasm by
using host cell enzymes
Then the proteins migrate into the nucleus and are oined !ith the
ne!ly synthesized DNA to form virions
These virions are transported along the endoplasmic reticulum to the
host cell"s membrane for release
#erpesviruses, papovaviruses, adenoviruses, and hepadnaviruses

all follo! this pattern of biosynthesis
$o%viruses& e%ception as all of their components are synthesized in

the cytoplasm
'ollo!ing attachment, entry, and uncoating, the viral DNA is

released into the nucleus of the host cell
Transcription and translation of a portion of the viral DNA- the &earl%

genes'
enzymes that are re(uired for the multiplication of viral DNA
)n most DNA viruses, early transcription is carried out !ith the host"s
transcriptase *+NA polymerase,
$o%viruses contain their o!n transcriptase
DNA replication occurs
transcription and translation of the remaining -late- viral genes
capsid and other structural proteins
capsid proteins migrate into the nucleus, host cell, maturation, the
viral DNA and capsid proteins assemble to form complete viruses
and release
(rowing Bacteriophages in the )aborator%
can be gro!n either in suspensions of bacteria in
li(uid media
bacterial cultures on solid media

the plaque method
A sample of bacteriophage is mi%ed !ith host bacteria and melted agar
The agar containing the bacteriophages and host bacteria is then poured
into a $etri plate
containing a hardened layer of agar gro!th medium
The virus-bacteria mi%ture solidifies into a thin top layer that contains a
layer of bacteria appro%imately one cell thic.
/ach virus infects a bacterium, multiplies, and releases several hundred
ne! viruses
'ollo!ing several multiplication cycles, all the bacteria in the area
surrounding the original virus are destroyed
This produces a number of clearings, or pla(ues, visible against a la!n
of bacterial gro!th
/ach pla(ue theoretically corresponds to a single virus
the concentrations of viral suspensions measured by the number of pla(ues
usually given in terms of pla(ue-forming units *$'0,1
(rowing Animal "ir!ses in the laborator%
three methods are commonly used

213iving animals&
41embryonated eggs
51cell cultures
)i#ing Animals
mice, rabbits, and guinea pigs
to study the immune system"s response to viral infections
as a diagnostic procedure for identifying and isolating a
virus from a clinical specimen
After the animal is inoculated !ith the specimen, the animal is

observed for signs of disease
.illed to e%amine the infected tissues

*mbr%onated *ggs
convenient and ine%pensive
A hole is drilled in the shell of the embryonated egg
a viral suspension or suspected vi rus-containing tissue is inected

into the fluid of the egg
there are several membranes in an egg, and the virus is inected

near the one most appropriate for its gro!th
6iral gro!th is signaled by

the death of the embryo
embryo cell damage
the formation of typical poc.s or lesions on the egg membranes
Still used to gro! viruses for some vaccines

Cell C!lt!res
have replaced embryonated eggs as the preferred type of gro!th

medium for many vi ruses
consist of cells gro!n in culture media in the laboratory
Generally homogeneous collections of cells
can be propagated and handled much li.e bacterial cultures
more convenient to !or. !ith than !hole animals or embryonated

eggs
started by treating a slice of animal tissue !ith enzymes that

separate the individual cells
These cells are suspended in a solution that provides the osmotic
pressure, nutrients, and gro!th factors needed for the cells to
Gro!
7ells adhere to the glass or plastic container and reproduce to form
a monolayer
6iruses infecting such a monolayer sometimes cause the cells of

the monolayer to deteriorate as they multiply
This cell deterioration is called cytopathic effect *7$/,
7$/ can be detected and counted

7ell lines are of different types&
Primar% cell lines:
derived from tissue slices
tend to die out after only a fe! generations
Diploid cell lines:

developed from human embryos
can be maintained for about 288 generations

!idely used for culturing viruses that re(uire a human host
used to culture rabies virus for a rabies vaccine
Contin!o!s cell lines (immortal cell lines)
transformed *cancerous, cells that can be maintained through an

indefinite number of generations
/%ample& the #e3a cell line
)solated from the cancer of a !oman *#enrietta 3ac.s, !ho died in

29:2
there are still some viruses that have never been successfully
cultivated in cell culture
Prions
proteinaceous infectious particle
%roposed as an infectius particle by !merican neurobiologist

,tanley %rusiner in +?@).
caused a neurological disease in sheep called scrapie
ine animal diseases caused by %rions have been identified
AMad cow diseaseB in cattle in /reat 'ritain in +?@C
!ll nine are neurological diseases
large vacuoles develop in the brain

Human diseases &y Prion
Duru
Creu tzfeldt1 Ea:ob disease (CJD)
/erstmann1,triiussler1,chein:er syndrome
&atal familial insomnia
4hese diseases run in families. indicating a possible genetic

cause
not purely inherited
Mad cow disease developed from feeding scrapie1infected

sheep meat to cattle
CED has been transmitted with transplanted nerve
t issue and contaminated surgical instruments-

Viroids
,hort pieces of na:ed "!
Only 9** to F** nucleotides long
o protein coat
4he nucleotides arc often internally paired
#ave a closed. folded. three1dimensional structure that

protect it from attac: by cellular enzymes
4he "! does not code for any proteins
(iroids have been identified as pathogens only of plants
Example5 potato spindle tuber viroid

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Copyright 2004 Pearson Education, Inc.

, publishing as Benjamin Cummings

Obligatory intracellular parasites

Multiply inside living cells by using the synthesizing

Exist as non living entity outside living cell

Cause the synthesis of specialized structures that can transfer

Most viruses infect only specific types of cells in one host

'acteria (Bacteriophages, or phages)

(iruses range from )* to +*** nm in length

Can be seen only by electron microscope

most are smaller than bacteria

,ome are almost similar to bacteria

Eg-. vaccinia virus

a complete. fully developed. infectious viral particle

composed of nucleic acid and surrounded by a protein coat

single or double stranded

%rotects nucleic acid from environment

(ehicle of transmission from one to another host

Each capsid is composed of protein subunits called

single or several types of protein

$n some viruses. the capsid is covered by an envelope

consists of lipid. protein and carbohydrates

0ayer of host cell plasma membrane for animal virus

$n some cases. envelop proteins determined by viral nucleic

pro2ect from the surface

3sed for attachment by some

3sed for identification

different morphological types on the basis of their capsid

long rods that may be rigid or flexible

4he viral nucleic acid is found within a

Example5 "abies virus

capsid of most polyhedral viruses is

a regular polyhedron with )*

4he capsomeres of each face

7hen helical or polyhedral viruses are

enveloped helical virus is the influenza virus

enveloped polyhedral (icosahedral) virus is the herpes

(iruses are classified by differences in the nucleic !cid

&amily names end in 1viridae

/enus names end in 1virus

! group of viruses sharing the same genetic information and

,ubspecies are designated by a number

Species: #uman herpes virus

Subspecies: ##(+. ##( ). ##( 9

4he basic mechanism is similar for all virus

Entry and exit might vary

4o multiply virus needs to infect a host and use its

Most enzymes are obtained from the host

&rom a single virion several or even thousands of similar

'acteriophages can multiply by two alternative mechanisms5

!ttachment %hage attaches by tail fibers to

%enetration %hage lysozyme opens cell wall.

'iosynthesis %roduction of phage D!

Maturation !ssembly of phage particles

"elease %hage lysozyme brea:s cell wall

&ollowed by chance collision

specific association;interaction of virion with specific receptors

'acterial mutant unable to synthesis specific receptors resistant

susceptible cell contain ca- +*

!bsorption highly specific for animal = bacterial virus but

(irus might enter via phagocytosis or some other endocytic

entrance of viral D! into host cell cytoplasm

$n bacteriophages. more complicated than animal = plant viruses