Virology

Virology




Mrs. Rebecca Thombre
Dept of Biotechnology
Modern College,
Shivajinagar,Pune-5
Historical

• Harmful agents were called viruses (Latin
  virus, poison or venom ) in the nineteenth
  century
• Louis Pasteur used the term virus to
  describe any living disease causing agent

• In 1892 ,Dimitri Ivanowski gave the first
  clear evidence of viruses.He observed that
  leaf extract of infected plant could induce
  tobacco mosaic disease.
• Beijerinck performed experiments on the
  same line and proposed that the disease is
  caused by a contagium vivum fluidum.

• During the same time Loeffler and Frosch
  observed that the foot and mouth disease
  of cattle was caused by a filterable virus.
• In 1911, Peyton Rous reported that virus was
  responsible for malignant muscle tumor in chicken.
• French scientist d’Herelle discovered bacterial
  viruses.
• Wendell Stanley crystallized tobacco mosaic virus in
  1935 and found that it was largely protein. Bawden
  and Pirie separated TMV particle into protein and
  nucleic acid. It is now clear that viruses are
  complexes of proteins and nucleic acids capable of
  replication only in living cells
Occurrence
• Viruses are unique infectious agents which
  reproduce only in living cells.
• They occur in bacteria,algae ,protozoa and
  higher plants and animals
General Properties

• Viruses are much smaller than bacteria.Their size
  ranges from 20 to 300 nm. Most viruses are invisible
  under the light microscope.
• They have very simple structure. They are
  nucleoprotein particles consisting of DNA or RNA
  surrounded by a protein capsid.
• They lack cellular organization.Cytoplasm,
  mitochondria, Golgi body ,lysososomes, cell membrane
  and ribosome are absent. They use ribosomes of host
  during protein synthesis.
• Viruses cannot multiply outside a living cell. They are
  obligate parasites.
• They are metabolically inactive. They lack enzyme
  systems and protein synthesis machinery.
• Generally cell have both DNA and RNA.They have
  only one nucleic acid, either DNA or RNA.
• They can be crystallized.
• Viruses donot have the power of growth and
  division.A fully formed virus cannot increase in
  size neither can it undergo division.
Structure
• The intact virus unit or infectious particle is called
  the virion. Each virion consists of a nucleic acid core
  surrounded by a protein coat called capsid.




• It is composed of a number of subunits called
  capsomeres. The nucleic acid and capsid is called
  nucleocapsid. The nucleocapsid may be naked or
  surrounded by a loose membrane called envelope.
Morphology
Viruses occur in three shapes: spherical, helical and complex




    HIV                     TMV               POX VIRUS
Icosahedral Symmetry
Virology
• Spherical symmetry (Polyhedral): Crick and
  Watson have shown that polyhedral capsids can
  have three possible types of symmetry viz
  tetrahedral, octahedral and icosahedral.
• Icosahedron is the most efficient shape for the
  packaging and bonding of subunits.
• An icosahedron is a regular polyhedron with 20
  faces formed by equilateral triangles and 12
  intersecting points or corners. Each capsids is
  made up of capsomeres.
• There are two types of capsomeres, pentameres
  and hexameres. Polyhedral capsids may be
  – Naked : Example Poliovirus, Adeno virus and
    papilloma virus or
  – Enveloped: Example Herpes Simplex virus
Enveloped with helical nucleocapsid (influenza virus)
Enveloped with icosahedral
nucleocapsid (herpesvirus)
Spherical Symmetry
Human Immunodeficiency Virus
Helical Symmetry

            Helical capsid consists of
            monomers arranged in a
            helix around a single
            rotational axis.
            The monomers curve into a
            helix because they are
            thicker at one end than the
            other.
            Helical capsids may be
            naked ,example
            bacteriophage M13
            ,tobacco Mosaic Virus
            (TMV)
• Helical symmetry (cylindrical): Helical capsid
  consists of monomers arranged in a helix
  around a single rotational axis.The monomers
  curve into a helix because they are thicker at
  one end than the other.
• Helical capsids may be naked ,example
  bacteriophage M13 ,tobacco Mosaic Virus
  (TMV) or
• enveloped ,example influenza virus.
Virology
• Complex symmetry : Complex viruses are
  those which are divided into two groups :
• a)Viruses without identifiable capsids .
e.g vaccinia virus



b) Viruses whose capsids are
attached with additional
structures.
e.g Bacteriophages of
T even series (T2,T4,T6).
• Envelope: Many viruses are surrounded by a 100
  – 150 A thick membrane called envelope. Virus
  envelope contains host cell proteins and proteins
  specified by virus. Some members have
  glycoprotein spikes. The lipids envelopes of
  budding viruses are derived from host.
• Nucleic acids: Viruses contain DNA or RNA, which
  may be single stranded (ss), double stranded (ds),
  linear or circular. The may have plus polarity or
  minus polarity
Viral Proteins
• Structural (nucleocapsid) proteins: Viral capsids are made up
  entirely of proteins. The capsid is made up of identical protein
  subunits called protomers. Some viral capsids have more than
  one type of protein. Capsid protein protects the viral nucleic
  acid from host nucleases and also helps in attachment to
  susceptible sites.
• Core (Internal) proteins: They are the proteins associated with
  nucleic acid of virion. E.g. Protein V and VII of Adeno virus and
  nucleoproteins of vesicular stomatitis virus and influenza virus.
• Viral Enzymes: Several enzymes are found in animal viruses.
  The ds RNA viruses contain enzymes for synthesis of viral
  mRNA. The major types of enzymes affect host cell surface, add
  specific terminal groups to viral mRNA, transcribe DNA to
  mRNA or participate in nucleic acid replication and processing.
• Envelope proteins: Viral envelopes contain host cell proteins as
  well as proteins specified by virus. Some envelopes also contain
  glycoprotein and lipoprotein.
TMV

• Tobacco mosaic virus : TMV is a rod shaped,
    helical virus about 300 nm long and 15 – 18 nm in
  diameter.It is made of 2130 identical protein subunits
  which are arranged around a central hole of 4 nm
  diameter.
• Genome :The TMV RNA genome is single stranded and
  linear, with a length of ~6400 bases. The tightly
  organized genome encodes at least three nonstructural
  proteins (P126, P183, and the 30-kD MP). Both P126
  and P183 are components of the TMV replicase. The
  genome also codes for a putative 54-kD protein of
  unknown function, and the Coat Protein.
Life cycle
• 1)Entry : The virus enters the plant cell through stomatal openings
  or through through mechanical injuries on the plant surface.
• 2) Replication : The virus passes from one cell to another via the
  plamodesmata. Several particles invade the nucleus. Replication
  takes place in the nucleus.
• 3) Synthesis of viral genome and proteins: Synthesis of RNA
  dependant RNA occurs in the nucleus. The capsid proteins are also
  synthesized in the nucleus.
• 4)Assembly : The assembly of protein subunits and nucleic acid
  starts within the nucleus.However incomplete viral particles may
  also come out of the cytoplasm and assembly may be completed
  there.After assembly the viral particles are released in the plant
  cell.
HIV
• HIV : Human Immunodeficiency Virus (HIV. is a retro virus
  which causes AIDS (Acquired Immune Deficiency
  Syndrome.) The immune deficiency is caused by the loss of
  the CD4+ T cells that are essential for both cell mediated and
  antibody-mediated immunity.HIV is spherical enveloped
  virus about 90 to 120 nm in size. The nucleocapsid has an
  outer icosahedral shell and an inner cone shaped core
  enclosing ribonucleoproteins. The genome is diploid
  composed of two identical single stranded positive RNA
  copies associated with viral reverse transcriptase enzyme.
• The virion binds to both CD4 and either coreceptor with
  molecules on its surface called glycoprotein 120 (gp120).
• This binding triggers an allosteric change in a second
  molecule, called glycoprotein 41 (gp41), which penetrates
  the host plasma membrane allowing the virion to get inside.
• When HIV infects a cell
• its molecules of reverse transcriptase and integrase are carried into
  the cell attached to the viral RNA molecules.
• The reverse transcriptase synthesizes DNA copies of the RNA.
• These enter the nucleus where the integrase catalyzes their
  insertion into the DNA of the host's chromosomes.
• The HIV DNA is transcribed into fresh RNA molecules which reenter
  the cytosol where
    – some are translated by host ribosomes.
        • The env gene is translated into molecules of the envelope protein
          (gp160).
          Proteases of the host cell then cut gp160 into
             – gp120 which sits on the surface of the virions (and is the target of
                most of the vaccines currently being tested).
             – gp41, a transmembrane protein associated with gp120.
        • the gag and pol genes are translated into a single protein molecule which
          is cleaved by the viral protease into
             – 6 different capsid proteins
             – the protease
             – reverse transcriptase
             – the integrase
• other RNA molecules become incorporated into fresh virus particles
Virology
T4
• T4 is a coliphage belonging to the T even series. It has a
  head, head tail connector, tail, baseplate and tail fibers
• The head capsid is a 95 nm long and 65 nm wide
  prolate icosahedron. It has DNA associated with several
  peptides and three internal proteins.
• The DNA is linear and has a molecular weight of 120 X
  106 .The head tail connector has a collar with attached
  whiskers.
• The tail has a contractile sheath surrounding an inner
  core or tube. It is connected to the collar at the upper
  end and baseplate at the lower end.
• The baseplate is hexagonal and has tail fibers attached
  to it.
T4 phage
Virology
Virology
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Virology

  • 1. Virology Mrs. Rebecca Thombre Dept of Biotechnology Modern College, Shivajinagar,Pune-5
  • 2. Historical • Harmful agents were called viruses (Latin virus, poison or venom ) in the nineteenth century • Louis Pasteur used the term virus to describe any living disease causing agent • In 1892 ,Dimitri Ivanowski gave the first clear evidence of viruses.He observed that leaf extract of infected plant could induce tobacco mosaic disease.
  • 3. • Beijerinck performed experiments on the same line and proposed that the disease is caused by a contagium vivum fluidum. • During the same time Loeffler and Frosch observed that the foot and mouth disease of cattle was caused by a filterable virus.
  • 4. • In 1911, Peyton Rous reported that virus was responsible for malignant muscle tumor in chicken. • French scientist d’Herelle discovered bacterial viruses. • Wendell Stanley crystallized tobacco mosaic virus in 1935 and found that it was largely protein. Bawden and Pirie separated TMV particle into protein and nucleic acid. It is now clear that viruses are complexes of proteins and nucleic acids capable of replication only in living cells
  • 5. Occurrence • Viruses are unique infectious agents which reproduce only in living cells. • They occur in bacteria,algae ,protozoa and higher plants and animals
  • 6. General Properties • Viruses are much smaller than bacteria.Their size ranges from 20 to 300 nm. Most viruses are invisible under the light microscope. • They have very simple structure. They are nucleoprotein particles consisting of DNA or RNA surrounded by a protein capsid. • They lack cellular organization.Cytoplasm, mitochondria, Golgi body ,lysososomes, cell membrane and ribosome are absent. They use ribosomes of host during protein synthesis. • Viruses cannot multiply outside a living cell. They are obligate parasites.
  • 7. • They are metabolically inactive. They lack enzyme systems and protein synthesis machinery. • Generally cell have both DNA and RNA.They have only one nucleic acid, either DNA or RNA. • They can be crystallized. • Viruses donot have the power of growth and division.A fully formed virus cannot increase in size neither can it undergo division.
  • 8. Structure • The intact virus unit or infectious particle is called the virion. Each virion consists of a nucleic acid core surrounded by a protein coat called capsid. • It is composed of a number of subunits called capsomeres. The nucleic acid and capsid is called nucleocapsid. The nucleocapsid may be naked or surrounded by a loose membrane called envelope.
  • 9. Morphology Viruses occur in three shapes: spherical, helical and complex HIV TMV POX VIRUS
  • 12. • Spherical symmetry (Polyhedral): Crick and Watson have shown that polyhedral capsids can have three possible types of symmetry viz tetrahedral, octahedral and icosahedral. • Icosahedron is the most efficient shape for the packaging and bonding of subunits. • An icosahedron is a regular polyhedron with 20 faces formed by equilateral triangles and 12 intersecting points or corners. Each capsids is made up of capsomeres. • There are two types of capsomeres, pentameres and hexameres. Polyhedral capsids may be – Naked : Example Poliovirus, Adeno virus and papilloma virus or – Enveloped: Example Herpes Simplex virus
  • 13. Enveloped with helical nucleocapsid (influenza virus)
  • 16. Helical Symmetry Helical capsid consists of monomers arranged in a helix around a single rotational axis. The monomers curve into a helix because they are thicker at one end than the other. Helical capsids may be naked ,example bacteriophage M13 ,tobacco Mosaic Virus (TMV)
  • 17. • Helical symmetry (cylindrical): Helical capsid consists of monomers arranged in a helix around a single rotational axis.The monomers curve into a helix because they are thicker at one end than the other. • Helical capsids may be naked ,example bacteriophage M13 ,tobacco Mosaic Virus (TMV) or • enveloped ,example influenza virus.
  • 19. • Complex symmetry : Complex viruses are those which are divided into two groups : • a)Viruses without identifiable capsids . e.g vaccinia virus b) Viruses whose capsids are attached with additional structures. e.g Bacteriophages of T even series (T2,T4,T6).
  • 20. • Envelope: Many viruses are surrounded by a 100 – 150 A thick membrane called envelope. Virus envelope contains host cell proteins and proteins specified by virus. Some members have glycoprotein spikes. The lipids envelopes of budding viruses are derived from host. • Nucleic acids: Viruses contain DNA or RNA, which may be single stranded (ss), double stranded (ds), linear or circular. The may have plus polarity or minus polarity
  • 21. Viral Proteins • Structural (nucleocapsid) proteins: Viral capsids are made up entirely of proteins. The capsid is made up of identical protein subunits called protomers. Some viral capsids have more than one type of protein. Capsid protein protects the viral nucleic acid from host nucleases and also helps in attachment to susceptible sites. • Core (Internal) proteins: They are the proteins associated with nucleic acid of virion. E.g. Protein V and VII of Adeno virus and nucleoproteins of vesicular stomatitis virus and influenza virus. • Viral Enzymes: Several enzymes are found in animal viruses. The ds RNA viruses contain enzymes for synthesis of viral mRNA. The major types of enzymes affect host cell surface, add specific terminal groups to viral mRNA, transcribe DNA to mRNA or participate in nucleic acid replication and processing. • Envelope proteins: Viral envelopes contain host cell proteins as well as proteins specified by virus. Some envelopes also contain glycoprotein and lipoprotein.
  • 22. TMV • Tobacco mosaic virus : TMV is a rod shaped, helical virus about 300 nm long and 15 – 18 nm in diameter.It is made of 2130 identical protein subunits which are arranged around a central hole of 4 nm diameter. • Genome :The TMV RNA genome is single stranded and linear, with a length of ~6400 bases. The tightly organized genome encodes at least three nonstructural proteins (P126, P183, and the 30-kD MP). Both P126 and P183 are components of the TMV replicase. The genome also codes for a putative 54-kD protein of unknown function, and the Coat Protein.
  • 23. Life cycle • 1)Entry : The virus enters the plant cell through stomatal openings or through through mechanical injuries on the plant surface. • 2) Replication : The virus passes from one cell to another via the plamodesmata. Several particles invade the nucleus. Replication takes place in the nucleus. • 3) Synthesis of viral genome and proteins: Synthesis of RNA dependant RNA occurs in the nucleus. The capsid proteins are also synthesized in the nucleus. • 4)Assembly : The assembly of protein subunits and nucleic acid starts within the nucleus.However incomplete viral particles may also come out of the cytoplasm and assembly may be completed there.After assembly the viral particles are released in the plant cell.
  • 24. HIV • HIV : Human Immunodeficiency Virus (HIV. is a retro virus which causes AIDS (Acquired Immune Deficiency Syndrome.) The immune deficiency is caused by the loss of the CD4+ T cells that are essential for both cell mediated and antibody-mediated immunity.HIV is spherical enveloped virus about 90 to 120 nm in size. The nucleocapsid has an outer icosahedral shell and an inner cone shaped core enclosing ribonucleoproteins. The genome is diploid composed of two identical single stranded positive RNA copies associated with viral reverse transcriptase enzyme. • The virion binds to both CD4 and either coreceptor with molecules on its surface called glycoprotein 120 (gp120). • This binding triggers an allosteric change in a second molecule, called glycoprotein 41 (gp41), which penetrates the host plasma membrane allowing the virion to get inside.
  • 25. • When HIV infects a cell • its molecules of reverse transcriptase and integrase are carried into the cell attached to the viral RNA molecules. • The reverse transcriptase synthesizes DNA copies of the RNA. • These enter the nucleus where the integrase catalyzes their insertion into the DNA of the host's chromosomes. • The HIV DNA is transcribed into fresh RNA molecules which reenter the cytosol where – some are translated by host ribosomes. • The env gene is translated into molecules of the envelope protein (gp160). Proteases of the host cell then cut gp160 into – gp120 which sits on the surface of the virions (and is the target of most of the vaccines currently being tested). – gp41, a transmembrane protein associated with gp120. • the gag and pol genes are translated into a single protein molecule which is cleaved by the viral protease into – 6 different capsid proteins – the protease – reverse transcriptase – the integrase • other RNA molecules become incorporated into fresh virus particles
  • 27. T4 • T4 is a coliphage belonging to the T even series. It has a head, head tail connector, tail, baseplate and tail fibers • The head capsid is a 95 nm long and 65 nm wide prolate icosahedron. It has DNA associated with several peptides and three internal proteins. • The DNA is linear and has a molecular weight of 120 X 106 .The head tail connector has a collar with attached whiskers. • The tail has a contractile sheath surrounding an inner core or tube. It is connected to the collar at the upper end and baseplate at the lower end. • The baseplate is hexagonal and has tail fibers attached to it.