CHAPTER 4
VIRUS ENTRY
STEPS IN VIRUS ENTRY
- How do virions get into cells
o Viruses of bacteria, archaea, algae and plants use
different entry mechanisms than anima viruses
o Bacteriophages may puncture through cell wall
o Plant viruses may enter through mechanical
means then spread through plasmodesmata
- Enveloped and nonenveloped viruses have distinct
penetration strategies
- Enveloped viruses can enter by:
o Fusion and Fission of the envelope with the
plasma membrane
o Receptor-mediated endocytosis followed by
fusion/fission with an endosome
- Entry of enveloped viruses
- Non enveloped viruses can enter by:
o Receptor-mediated endocytosis
o Passage of genome through a channel in the
membrane
- Some viruses can pass directly from cell to cell
o Virus passed to neighboring cell by filopodia
o Virus assembled and transmitted at virological
synapse
o Formation of syncytia
- A variety of cell surface proteins can serve as specific virus
receptors or attachment factors
- Receptors interact with viral glycoproteins, surface
protrusions, or canyons
- Many viruses enter the cell via receptor-mediated
endocytosis
o May occur at clathrin-coated pits, caveolae or
lipid rafts
o Virus then delivered to early endosomes
- Some viruses enter by macropinocytosis
PENETRATION THROUGH CELLULAR MEMBRANES
- Passage from endosomes to the cytosol is often triggered
by a low pH
o Induces conformational changes in virus
- Membrane fusion is mediated by specific viral fusion
proteins
o Generally type I transmembrane proteins that
contain fusion peptides
o Class I fusion proteins
o Class II fusion proteins
- Fusion proteins undergo major conformational changes
that lead to fusion
o Activated by low pH or receptor binding
- Non enveloped viruses penetrate by membrane lysis or
pore formation
o Genomes may pass through pores formed by the
virus
o Some viruses rupture endosomal membrane
INTRACELLULAR TRANSPORT
- Virions and capsids are transported within the cell in
vesicles or on microtubules
o Dyenin and dynactin move viruses along
microtubules
- Import of viral genomes into the nucleus
o Interaction with importins, karyopherins
o Dissociation of the nuclear membrane during
mitosis
o Import after disassembly at the nuclear pore
o Transport of intact virions through the nuclear
port
- The many ways in which viral genomes are uncoated and
released
o May occur simultaneously with entry
o May occur after a complex serious fusion of
transport steps
HOW CAN VIRUS ENTRY BE PREVENTED?
- Intercept virus before it reaches cell with neutralizing
antibodies
- Flood extracellular space with soluble receptor
- Block cellular receptor
- Lysosomotropic agents, carboxylic ionophores and
bafilomycin A1 inhibit acidification of endosome
- Inhibit membrane fusion
- Inhibit uncoating (amantadine, WIN compounds)
FUNDAMENTAL CONCEPTS
 - Viral proteins bind to specific receptors on cellular surface c. After uncoating, the infectious particles cannot
- Entry may occur by fusion, endocytosis or passage of viral be detected
genome through a pore 4. Synthesis
- Fusion may be mediated by low pH - TO create new NA, need polymerases
- Nucleocapsids can be transported on microtubules o IF DNA
- Uncoating of genomes can occur by a variety of means DNA polymerases
o IF RNA
RNA polymerases

In general
First synthesized
1. Enzymes necessary for replication of nucleic acids
(Polymerases)
2. Regulatory proteins
Later synthesized
1. Protein necessary to construct progeny virions
2.
I. Replication of the viral nucleic acid can be in the
cytoplasm or the nucleus
- Most of the DNA viruses sa nucleus nagssynthesize
- Most of the RNA viruses sa cytoplasm nagssynthesize
II. Synthesis of the viral protein is exclusively in the
CYTOPLASM
III.
b. Host Mediated
c. Virus-Mediated
A. DNA viruses
- Diffuse into nucleus
- Transcription is initiated by cellular DNA-dependent RNA
polymerases
- Only a part of viral genome is transcribed and translated
1. Lead to production of early proteins (Matrix proteins,
Regulatory proteins, Virally encoded polymerases)
2. Production of late proteins (Structural proteins needed to
assemble the progeny virions, capsid proteins)

B. +RNA
i. Early period
Viral-RNA dependent
RNA-polymerase, a protease and several structural
proteins are synthesized
Common Stages of Viral Replicative Cycles ii. New polymerase copies +RN) strands and converts to
RNA strands
1. Adsorption / Attachment Serve as templates for the synthesis of a large number
a. Initial stage of infectious cycle of +RNA strands ----viral progeny virions
b. Adherence to a specific receptor
i. Multiplicity of infection C. RNA
2. Penetration / Injection (phages) - Not infectious
a. Membrane fusion (enveloped viruses) - Need to be transcribed into +RNA strands before viral
b. Receptor mediated endocytosis (non-enveloped proteins can be synthesized
viruses) o Pre-packed with virally encoded RNA polymerase
3. Uncoating of viral nucleic acid - Host cells do not have RNA dependent RNA polymerase
a. Nucleocapsids migrate in the cytoplasm / nucleus i. Early period
i. Non enveloped virus Synthesis and transcription of +RNA
1. Direct when penetration is It will produce
successful > short RNA transcript
ii. Enveloped virus > long RNA transcript
1. When receptor mediated ii. Late period
endocytosis is the mechanism 1. Short + RNA strands
of entry, acid H in the Translated into several viral proteins and
endosome promotes fusion enzymes to form new viral progeny
of viral envelope and 2. Long + RNA strands
endosomal membrane Serve as template for progeny
b. Cytoplasmic proteases/ Viral proteases digest -RNA strands accumulation will end up with viral
protein layer (capsid) polymerases

IV. Condensation
1. Viruses with small genomes are more restricted
2. In general, the bigger the viral genome, the more complex
the structure can be
3. Viruses with larger genomes provide more of the functions
they need to replicate and rely less on the production of
specific enzymes by the cell
4. With small genomes, a single protein frequently has
multiple functions
5. Many viruses with small size genomes, use methods of
making one genome sequences produce multiple proteins
and common ability to self-assemble -> overlapping genes
Morphogenesis / Assembly
Book-Based
Virus Classification: The World of Viruses
STEPS IN VIRUS ENTRY
- Virions bind to specific receptors on the cell surface
- Virions enter the cell either in specialized vesicles or by
direct penetration through the plasma membrane
- Virions or nucleocapsids are transported within the cell to
sites of transcription and replication (cytoplasm or nucleus)
- Viral genomes are released into the cell by passage through
the vesicle membrane or by disintegration of capsids within
the cell (uncoating)
PENETRATION THROUGH CELLULAR MEMBRANES
- Membrane penetration is mediated by conformational
changes of virion proteins
- These changes are triggered by
o Interaction of viral surface proteins with cellular
receptors in membranes or
o Exposure to low pH inside endosomes
- Enveloped viruses penetrate by fusion of the virus envelope
with either:
o Plasma membrane or
o Membranes of intracellular vesicles
- Non-enveloped viruses enter the cytosol by either
o Forming transmembrane channels through which
the viral nucleocapsid or genome can pass, or
o Inducing lysis of endocytic vesicles
HOW DO VIRIONS GET INTO CELLS?
- The transfer process begins with the assembly of virions in
the infected cell
o Release
Transmission (organism or
environment)
Entry (to a new host cell
- Viruses follow the molecular rules and regulations that
control events inside the cell, but manipulate these
pathways to serve their own ends
- Enter plant through intercellular channels called
plasmodesmata normally allow communication and
passage of metabolites
Enveloped and non-enveloped viruses have distinct penetration
strategies
- Enveloped viruses circumvent the problem by using vesicle
transport and couple fusion (coming together) and fission
(breaking apart) of membranes
o Viral envelope = transport vessel
o Nucleocapsid = cargo
- The viral envelope forms around the nucleocapsid and
other internal components such as matrix proteins during
budding at a cellular membrane .
- Budding
o Often at the PM (external)
o But some. Use the membranes of the nucleus, ER,
intermediate compartment, endosomes or Golgi
complex
- Delivery of the nucleocapsid cargo into the cytosol of the
target cell occurs when the viral envelope fuses with a
cellular membrane
- Fusion can take place either
o PM
o Membranes of early or organelles in which the
virion is enclosed after undergoing late
endosomes
- Endosomes are membranous organelles in which the virion
is enclosed after undergoing receptor mediated
endocytosis
- The membrane fusion pathway has the advantage that viral
macromolecules do not need to pass directly across the
hydrophobic interior of any cellular membrane
- Non-enveloped viruses
o To exit from infected cell, they typically rely on
rupture of cellular membranes
o Cell death is usually necessary to release virus
particles trapped in the nucleus or the cytoplasm
o Enter the host cell in variety of ways
Adenoviruses
Taken up into endosomes,
where they proceed to
rupture the endosomal
membrane, releasing the
virion and other endosomal
contents into the cytosol
Picornaviruses
Undergo conformational
changes during binding to
surface receptors or after
entering endosomes, so that
viral proteins form
membrane channels that
allow the genome direct
access to the cytosol
Some viruses can pass directly from cell to cell
- Particless still associated with the surface of the infected
cell may reach neighboring cells through contacts
established via filopodia and other actin-based extensions
- In such cases, the infected host cells play an active role in
transmitting the infection, with minimal exposure of the
virus to the extracellular environment
- This can also help to avoid recognition of the virus by the
host immune system.
- Some retroviruses have been shown to generate a special
region where viruses are preferentially assembled and
transmitted a virological synapse in the contact area
between the virus-producing cell and its neighbor
- In few cases, infection can spread from one cell to the next
without formation of a complete virus particle
- Measles and some other enveloped viruses induce fusion of
infected cells with neighboring uninfected cells
o Results in formation of multinucleated cells called
syncytia
o The same viral proteins that serve as