DENGUE VIRUS JM 130110110036 A3

DENGUE VIRUS
Dengue is caused by Dengue virus (DENV), a mosquito-borne flavivirus. DENV is
an single stranded RNA positive-strand virus of the family Flaviviridae, genus Flavivirus.
This genus includes also the West Nile virus, Tick-borne Encephalitis Virus, Yellow Fever
Virus, and several other viruses which may cause encephalitis. DENV causes a wide range
of diseases in humans, from a self limited Dengue Fever (DF) to a life-threatening
syndrome called Dengue Hemorrhagic Fever (DHF) or Dengue Shock Syndrome (DSS).
There are four antigenically different serotypes of the virus: DENV-1, DENV-2,
DENV-3, DENV-4
Here, a serotype is a group of viruses classified together based on their antigens on the surface of
the virus. These four subtypes are different strains of dengue virus that have 60-80% homology
between each other. The major difference for humans lies in subtle differences in the surface
proteins of the different dengue subtypes. Infection induces long-life protection against the infecting
serotype, but it gives only a short time cross protective immunity against the other types. The first
infection cause mostly minor disease, but secondary infections has been reported to cause severe
diseases (DHF or DSS) in both children and adults. This fenomenon is called Antibody-Dependent
Enhancement.
Antibody-Dependent Enhancement- After a person is infected with dengue, they develop an immune response to
that dengue subtype. The immune response produced specific antibodies to that subtype specific surface proteins that
prevents the virus from binding to macrophage cells (the target cell that dengue viruses infect) and gaining entry. However, if another subtype of dengue virus
infects the individual, the virus will activate the immune system to attack it as if it was the first subtype. The immune system is tricked because the four subtypes
have very similar surface antigens. The antibodies bind to the surface proteins but do not inactivate the virus. The immune response attracts numerous
macrophages, which the virus proceeds to infect because it has not been inactivated. This situation is referred to as Antibody-Dependent Enhancement (ADE) of
a viral infection. This makes the viral infection much more acute. The body releases cytokines that cause the endothelial tissue to become permeable which
results in Dengue Haemorrhagic Fever (DHF) and fluid loss from the blood vessels.
STRUCTURE

DENV is a 50-nm virus enveloped with a lipid membrane. There are 180 identical copies of the envelope (E) protein
attached to the surface of the viral membrane by a short transmembrane segment. The virus has a genome of about 11000
bases that encodes a single large polyprotein that is subsequently cleaved into several structural and non-structural mature
peptides. The polyprotein is divided into three structural proteins, C, prM, E; seven nonstructural proteins, NS1, NS2a, NS2b,
NS3, NS4a, NS4b, NS5; and short non-coding regions on both the 5' and 3' ends. The structural proteins are the capsid (C)
protein, the envelope (E) glycoprotein and the membrane (M) protein, itself derived by furine-mediated cleavage from a prM
precursor. The E glycoprotein is responsible for virion attachment to receptor and fusion of the virus envelope with the target
cell membrane and bears the virus neutralization epitopes. In addition to the E glycoprotein, only one other viral protein, NS1,
has been associated with a role in protective immunity. NS3 is a protease and a helicase, whereas NS5 is the RNA polymerase in
charge of viral RNA replication.
LIFE CYCLE
The life cycle of dengue involves endocytosis via a cell surface receptor. The virus uncoatsintracellularly via a specific
process. In the infectious form of the virus, the envelope protein lays flat on the surface of the virus, forming a smooth coat with
icosahedral symmetry. However, when the virus is carried into the cell and into lysozomes, the acidic environment causes the
protein to snap into a different shape, assembling into trimeric spike. Several hydrophobic amino acids at the tip of this spike
insert into the lysozomal membrane and cause the virus membrane to fuse with lysozome. This releases the RNA into the cell
and infection starts.
DENGUE VIRUS JM 130110110036 A3

The DENV RNA genome is in the infected cell translated
by the host ribosomes. The resulting polyprotein is subsequently
cleaved by cellular and viral proteases at specific recognition
sites. The viral nonstructural proteins use a negative-sense
intermediate to replicate the positive-sense RNA genome, which
then associates with capsid protein and is packaged into
individual virions. Replication of all positive-stranded RNA viruses
occurs in close association with virus-induced intracellular
membrane structures. DENV also induces such extensive
rearrangements of intracellular membranes, called replication
complex (RC). These RCs seem to contain viral proteins, viral RNA
and host cell factors. The subsequently formed immature virions
are assembled by budding of newly formed nucleocapsids into
the lumen of the endoplasmic reticulum (ER), thereby acquiring a
lipid bilayer envelope with the structural proteins prM and E. The
virions mature during transport through the acidic trans-Golgi
network, where the prM proteins stabilize the E proteins to
prevent conformational changes. Before release of the virions
from the host cell, the maturation process is completed when prM is cleaved into a soluble pr peptide and virion-associated M
by the cellular protease furin. Outside the cell, the virus particles encounter a neutral pH, which promotes dissociation of the pr
peptides from the virus particles and generates mature, infectious virions. At this point the cycle repeats itself.
BUILDING NEW VIRUSES
Dengue virus also makes several proteins that create new viruses
once it is inside a cell. Two of the major ones are shown here. Both are
multifunctional proteins with several enzymes strung together. The one on the
left, NS5 from PDB entries 1l9k and 2j7w, contains a methyltransferase and a
polymerase, and the one on the right, NS3 from PDB entry 2vbc, contains a
protease and a helicase. Each of these enzymes performs a different part of
the life cycle. The polymerase builds new RNA strands based on the viral RNA,
the helicase helps to separate these strands, and the methyltransferase adds
methyl groups to the end of them, protecting the RNA strands and coaxing the
cell's ribosomes to create viral proteins based on them. The viral proteins are
created in one long polyprotein chain, which is finally clipped into the
functional units by the protease. The little chain colored blue is a portion of
another viral protein, NS2B, that assists with the protease activity.
VACCINE RESEARCH
There is no vaccine to protect against dengue. Although progress is underway, developing a vaccine against the disease is
challenging. The problem is:
With four different serotypes of the dengue virus that can cause the disease, the vaccine must immunize against all four
types to be effective.
There is still limited knowledge of how the disease typically behaves and how the virus interacts with the immune
system.
Another difficulty is that there is no reliable animal model for DHF and thus also not a suitable animal model to test
immune responses to potential vaccines.
In addition, progress in vaccine development is slow mainly because dengue viruses grow poorly in cell culture

References :
http://www.denguevirusnet.com/dengue-virus.html
http://www.rcsb.org/pdb/101/motm.do?momID=103