VIRUS REPRODUCTION

A. T. Aman
DEPT. OF MICROBIOLOGY,
FAC. OF MEDICINE, UGM, YOGYAKARTA.
 VIRUS ?
*. Infectious agents.
*. Genetic Elemen (genome) : DNA/RNA.
- One molecule/ more ,
- Single stranded (ss): Post./ neg. sense
- Double stranded (ds): Full/ partial ds
- NO: combination DNA & RNA.
*. Two phases: intracellular & intracellular.
intracellular: Submicroscopic particle: virion.
*. No. Metabolism.
*. Intracellular: genome replication, synthesis of
viral components, product. of complete virion.
 Basic Sructure of Virion
.
.
 Basic Sructure of Virion
*. Size and component: Vary.
*. Genome:
-. Virus with envelope: 1-2 %
-. Virus without envelope: 25-50%
. -. Segmented, > 1 mol.
#. Influenza virus: 8 mol. RNA; Rotavirus: 11 mol. RNA
- Partial dsDNA /RNA: Hepatitis B virus: partial dsDNA
*. Capsid: capsomere, contains info. for self
assembly
*. Some, have enzymes:
- RNA/DNA pol., Reverse transcriptase.
*. Some, more complex: + envelope (lipids).
- e.g.: influenza viruses, bacteriophage:
Hepatitis virus.
PRINCIPAL EVENTS IN VIRUS
REPLICATION
 1. ABSORPTION

*. Attachment: specific binding of a viral attachment

protein (VAP) to a cellular receptor.

*. Receptor: proteins (usually glycoproteins ), or the

sugar residues present on glycoproteins or glycolipids
(less specific).

Example:HIV: Receptor CD4 and VAP: gp120

(glycoprotein).
 1. ABSORPTION
*. Complex viruses (e.g. Poxviruses, Herpesviruses)
may have more than one receptor..> alternative routes of
uptake into cells.

*. Specific receptor binding can be side-stepped by

antibody-coated virus particles binding to Fc receptor on
the surface of monocytes, which results in virus uptake.
 1 . ABSORPTION
*. The expression of receptors determines the TROPISM
(cell type in which they can replicate)

*. Important in pathogenesis.

*. Attachment is a reversible process - if penetration does

not ensue, the virus can elute from the cell surface.

*.Elution from cell may lead to changes in the virus which

decrease or eliminate the possibility of attaching to other
cells
 2. PENETRATION
*. It is an energy-dependent process, i.e. the cell must be
metabolically active.
*. 3 mechanisms:
Translocation across the cell membrane
ENDOCYTOSIS into intracellular vacuoles;
eventually into the cytoplasm.
FUSION of the envelope with the cell membrane.
Requires: a viral fusion protein in the virus envelope,
e.g. influenza: haemagglutinin; retrovirus: envelope
glycoprotein.
 3. UNCOATING

*. Events after penetration: the capsid is removed, the

virus genome exposed.
-. Picornaviruses (simple) have a small basic protein
(VpG) covalently attached to the 5' end of the vRNA
genome
-. Retrovirus (complex): complex cores, also contain
the RT.
-. The process: inside the core particle.
-. That replicate in cytoplasm (Picornaviruses): genome
is released into the cell.
 3. UNCOATING
*. That replicate in the nucleus, (e.g.
Herpesviruses), the genome, associated
nucleoproteins, transported through the nuclear
membrane.

*. Achieved by interactions of the nucleoproteins

(or capsid) with the cytoskeleton.

*. At the nuclear pores, the capsid is stripped off,

and the genome passes into the nucleus.
 4. GENOME REPLICATION
The replication strategy depends on the genome nature.
7 groups:
I: dsDNA (Adenoviruses; Herpesviruses; Poxviruses, )
II: ss (+)sense DNA (Parvoviruses)
III: dsRNA (Reoviruses; Birnaviruses)
IV: ss (+)sense RNA (Picornaviruses; Togaviruses)
V: ss(-)sense RNA (Orthomyxoviruses,
Rhabdoviruses)
VI: ss (+)sense RNA with DNA intermediate in life-
cycle (Retroviruses)
VII: ds DNA with RNA intermediate (Hepadnaviruses
 I. Double-stranded DNA

e.g: Adenoviruses; Herpesviruses; Poxviruses.

Two groups:
a) Replication is exclusively nuclear (e.g.
Adenoviruses, Papovaviruses, Herpesviruses).

b) Replication occurs in cytoplasm (e.g. Poxviruses).

-. evolved all the necessary factors for transcription/
replication..> more independent of the cellular
machinery
 II. ss(+)sense DNA

Replication occurs in the nucleus,

involving the formation of a (-)sense
strand, which serves as a template for
(+)strand synthesis.
 III. ds RNA (Reoviruses).

*. These viruses have segmented

genomes. Each genome segment is
transcribed separately to produce
monocistronic mRNAs
 IV. ss (+)sense RNA

E.g.:Picornaviruses; Caliciviruses; Togaviruses;

Flaviviruses; Coronaviruses):
Two groups:
a) Polycistronic mRNA e.g. Picornaviruses; Hepatitis A.
Genome RNA = mRNA. Translation results in the
formation of a polyprotein product, which is
subsequently cleaved to form the mature proteins.
b) Complex Transcription e.g. Togaviruses. Two or
more rounds of translation are necessary to produce
the genomic RNA.
 V. ss(-)sense RNA

Can be:
a) Segmented e.g. Orthomyxoviruses.
-. Step 1.transcription of the (-)sense RNA genome
by the virion RNA-dependent RNA polymerase to
produce monocistronic mRNAs, which also serve
as the template for genome replication.

b) Non-segmented e.g. Rhabdoviruses. Replication

occurs as above and monocistronic mRNAs are
produced.
 VI. ss (+)sense RNA with DNA
intermediate

*. E.g.: Retroviruses:
-. Genome is (+)sense, unique among
viruses,
-. Not serve as mRNA, but as a template
for reverse transcription.
 VII. dsDNA with RNA intermediate

*E.g.: Hepadnaviruses:
Use RT (unlike the Retroviruses), this occurs inside
the virus particle on maturation.
*. On infection, the first event is repair of the
gapped genome, followed by transcription.
*. To replicate its genome, the virus must present
mRNA to the cell, and translated into virus-
encoded proteins. This can be done in a number of
ways, e.g:
 VII. dsDNA with RNA intermediate

* Done in a number of ways:

d/s DNA Viruse release their genome in the
nucleus, usen transcription cellular machinery.
(+)sense RNA viruses: genomes serve as
mRNA (except retroviruses!)
(-)sense RNA viruses: must carry with them
virus-coded enzymes for RNA-dep. RNA
replication.
 5. GENE EXPRESSION
*. replication Control : regulation of gene
expression.
*. Methods: depend on genome nature
/replication strategy.
Segmented genomes: transcribed to produce
monocistronic mRNAs.
Advantage of monocistronic: various proteins
can be produced in different amounts.
Non-segmented: tend to produce polycistronic
mRNA, .> form a polyprotein,
 5. GENE EXPRESSION

*. Use cellular machin: viral mRNAs

contain control signals recognized host cell
(ribosome-binding sites, etc).

Some DNA viruses (e.g. Papovaviruses),

encode a protein, binds to the Ori of rep.,
stimulates cellular DNA pol. to replicate the
virus genome.
 5. GENE EXPRESSION

Some viruses, e.g. Adenoviruses, encode

DNA pol., dependent on other host factors.

"Complex" viruses, e.g. Herpesviruses,

encode more proteins for DNA synth.,.>
more independent host machinery.
 6. ASSEMBLY
*. Assembly of all the components to form
mature virions at a particular site in the cell.
*. The basic structure is formed.
*. The site varies for different viruses., e.g:
Cytoplasm: Picornaviruses, Poxviruses,
Reoviruses.
Nucleus: Adenoviruses, Papovaviruses,
Parvoviruses.
Inner surface of cell membrane: Retroviruses
 7. RELEASE
*. Lytic viruses (most non-enveloped viruses): simple
- cell breaks, open & releases the viruses.
*. Enveloped viruses acquire the lipid membrane as
the virus buds out through the cell membrane.
*. Virion envelope protein are picked up during
this process as the virus is extruded.
*. Budding may/ may not kill cell
*. Controlled by virus: interaction of the capsid
on the inner surface of the cell membrane.
 8. MATURATION
*. Virus becomes infectious.
*. Structural changes, resulting from specific
cleavage of capsid proteins to form mature
products, conformational change in the capsid,
or condensation of nucleoproteins with the genome.
*. Some: assembly & maturation are inseparable, *.
Others: maturation occur after the virus particle
left cell.
 Bacteriophage REPRODUCTION

1. Attachment (absorption): virion to a susceptible

host.
2. Penetration (injection) into the cell by the virion or
by its NA.
3. Early events of multiplication:
By host replication machinery.
Some: viral enzymes involve.
PHAGE INFECTION
 REPRODUCTION
(contd)
4. Replication of genome.
5. Synthesis of protein subunits of the virus
coat
6. Assembly of NA and components of protein
subunits (membrane component).> new
virion.
7. Release of mature virion from the cell
(some cell: lysis).
 VIRUS
REPLICATION
.