Platelet Destruction Virus: A Description of the Virus and how it Interacts with a Human Host

Kaitlyn Howden
1136043
Gabriela Chang, T03
Hth Sci 3K03

Introduction:
The Platelet Destruction Virus (PDV) is a novel virus that has a tropism for human
platelet cells. These are cells that are derived from megakaryocytes, which are cells that have
differentiated from hematopoietic stem cells in the bone marrow. Megakaryocytes are progenitor
cells in this tissue that break off into fragments, which are the platelets, and these then released
into circulation to join other cell types in the blood, such as red blood cells and lymphocytes1.
The have a shot life span of only 8-10 days, but one trillion platelets circulate in the blood of an
adult human, and 100 billion new platelets are made from the bone marrow daily1. They play a
major role in blood clotting, which is necessary to prevent bleeding during injury, and a function
that is recently being recognized in platelets is their ability to aid in both innate and adaptive
immune function1. Interestingly, even though platelets are enucleated cells, they still maintain the
ability to carry out protein synthesis, and thus contain entities such as mRNA and ribosomes2.
These critical cytoplasmic functions required for the platelet to perform its role in mammals are
what RNA viruses, such as PDV, take advantage of for replication and progeny production. By
manipulating these cells, replicating in them at a fast rate, and then leading to their destruction,
the virus can cause an acute infection that leads to high viremia in the blood and massive
amounts of bleeding because of decreased clotting ability. Then when an unsuspecting host is
subjected to an injury, or a vector such as a mosquito bites the individual, the virus has a high
chance of passing its progeny along to another host. The rest of this paper will be devoted to
discussing specific aspects of this virus, and then describing how this interaction with the virus
affects its host.
Virus Structure and Genome:
PDV is a non-enveloped virus with an icosahedral capsid that contains a positive stranded
RNA genome. This genome codes for 8 genes, which are then transcribed into one long open

reading frame (ORF) that forms one large poly-protein. When this poly-protein is cleaved and
processed by viral proteases that it encodes, 4 structural and 4 nonstructural proteins are
generated. The structural proteins include 4 capsid proteins, CP1, CP2, CP3, and CP4 that act in
a similar fashion to the capsid proteins that comprise the poliovirus. CP1-3 fit together to make a
trimer, and each of these trimers comes together to make the whole icosahedral capsid. The forth
capsid protein, CP4, is a monomer that works to help hold and stabilize the trimer pieces together
and locks them in position. CP1 is also important in viral entry, as will be described later. The
virus also encodes non-structural proteins that dont contribute to the viral particle architecture,
but play a role in genomic replication or in mediating viral-host interactions. A major nonstructural protein is the RNA dependent RNA polymerase (RP) that the virus requires to make
more copies of its genome. This is necessary because humans do not encode a polymerase that
can read RNA as a template3. While the virus need not bring this in its capsid when first infecting
a host cell, because positive stranded RNA can be read directly by a ribosome, the virus must
encode it so that it can make a negative stranded copy of its genome3. This can then be used as a
template for the enzyme to produce multiple copies of the positive strand that can be packaged
into future progeny. Another non-structural protein is the protease (P), which is required to
process the poly-protein generated by the one long ORF encoded by the virus, allowing the
individual proteins to be formed. In order for the virus to egress out of the cell, since it is a nonenveloped virus, it needs to include the platelet death protein (PDP) that induces apoptosis. The
virus also includes features that provide it ways to overcome its host, including a novel protein
called inhibitor of SNARE complex (ISC) and an IRES structure in its 5 un-translated region
(UTR). Both of these features, and how they allow the virus to thrive in the host, will be
discussed later.

Life Cycle:
This virus, because it is transferred via blood contact, either needs a nave host to come
directly into contact with the blood of an infected individual, or needs a vector to carry the virus
from the blood stream of one person to another. The first scenario can be accomplished either by
contacting contaminated blood directly, since an individual with this disease will be unable to
clot and will therefore be bleeding excessively4. Contamination by another persons blood can
also happen by tainted IV or organ transplant procedures. A more common cause than this
though will be an arthropod like mosquitos and ticks biting an individual infected with this
disease and passing it to another individual. Since PDV creates a high viremia in the blood, there
will be enough of a viral load picked up by the vector that can be dispensed in a new host. Once
this occurs, the virus will travel in the bloodstream until it comes into contact with a platelet. It
will bind to the aIIbb3integrinreceptor,whichisthemostabundantsurfaceexpressed

integrinonplatelets,wherethereare40,00080,000copiesperplatelet5.Thetypical
ligandsforthisreceptorincludefibrinogen,fibrin,andvonWillebrandfactor(VWF)5.
Then, the CP1 protein will undergo a conformational transition that leads to its insertion into the
membrane. This creates a pore through which the viral RNA genome can be released into the
cytoplasm of the platelet.
Once inside, the positive stranded RNA can be directly translated by the ribosome to
make a poly-protein that can be cleaved by a protease that PDV encodes for. It can excise itself
and then continue to cleave the rest of the poly-protein to make the individual protein segments.
By the action of the ribosome and the RNA dependent RNA polymerase, multiple copies of the
genome and the structural proteins can be generated. The CP 1-4 proteins can spontaneously
assemble together to make the capsid, and then can associate with the viral genome and make
sure it gets incorporated into the virion. As the virus accumulates, it needs a way to get out of the
4

cell to spread its progeny. The PDP protein coded for by the virus is able to trigger an intrinsic
pathway of apoptosis, where mitochondrial instability leads to activation of caspase proteins,
which lead to protein and DNA degradation, and ultimately cell lyses6. Platelet cells undergoing
apoptosis cause a huge burst of virus to be released into the blood stream. Tens of thousands of
viral progeny can be released from only a couple of hours of replication time in the host cell.
This is an important feature of the virus since the life span of the platelets is so short, and
because it is in competition with the defending immune system. Once the infected cell bursts and
creates a high viral load in the system, this allows PDV to be picked up by another platelet, by a
vector, or directly by another host through mechanisms described previously.
Viral/Host interactions:
There are many features of the non-structural proteins, or the replication style of the
virus, that aid in giving it the upper hand when interacting with its host. For instance, similar to
poliovirus, the platelet destruction virus contains an IRES structure at its 5 end that substitutes
the use of a cap7. Human mRNA is typically capped at the 5 end7. This is important not only for
the stability of mRNA, since they are by nature easily degraded, but also because the platelet
utilizes standard cellular recruitment mechanisms of the ribosome to the mRNA for translation7.
The cap binding protein eIF4E recognizes the cap and it recruits other proteins like eIF4G that
will ultimately lead to bringing the small and large subunit of the ribosome around the mRNA so
it can be transcribed8. However, the IRES occurs when the genome takes on a base paired stem
loop structure that resembles the pre-ribosomal translation complex7. This secondary structure
mimicry allows formation of the ribosomal translation machinery and skips the requirement for a
cap7. The viral protease it encodes can also recognize the eIF4G protein and cleave a portion of it
off that is necessary for eIF4E binding8. The end goal of all of this is so the virus can hoard the
translational machinery for itself, and prevent the host cell from making the proteins it needs.
5

While the virus can hijack protein production, the cell it infects will have proteins already
that it can use for both its clotting and immunological functions, and these proteins must also be
dealt with. A good portion of the receptors or soluble factors the platelet uses to perform its
clotting function or to interact with cells of the immune system exist in the granules of an
inactivated platelet, and then are recruited to the cell membrane or secreted when the platelet
becomes activated9. These factors include P-selectin, fibronectin, CD154, and various cytokines
and antimicrobial substances9. They play roles in platelets activation and recruitment to damaged
endothelial tissue, and can affect the platelets interactions with lymphocytes such as
macrophages, B cells, and T cells1,9. To get rid of these functions, PDV encodes a non-structural
protein called ISC that functions to inhibit the SNARE complex used during secretion and
exocytosis10. The complex at the plasma membrane responsible for exocytosis is comprised of
SNARE proteins embedded in the plasma membrane itself and SNARE proteins embedded in the
granules/vesicles that combine and interact in order to fuse the two membranes10. The ISC
protein encoded by the virus specifically binds to a portion of the granule bound SNARE protein
called VAMP8, preventing proper complex formation10,11. This is extremely detrimental to
normal platelet function, but has no negative effect on viral entry or egress because it is a nonenveloped virus that is not dependent on exocytosis in order to get out of the cell.
While PDV doesnt specifically code for genes that aid it in avoiding immune detection,
RNA viruses are able to outcompete the immune system by their replication style. They replicate
on membrane surfaces in the cell, which is beneficial in this circumstance because platelets not
only have membrane-bound organelles like the ER, but also contains the membrane bound
granules that are trapped because of the actions of the ISC protein12. These surfaces are where
enzymes like RNA polymerase congregate into lattices and make transcription and replication

factories that are quick to make many copies of the viral genome12. However, two features of this
process create changes in the genome over time. First, RNA polymerases carried by this virus
have low fidelity, and often add mutations or mistakes into the genome13. Second, strand
switching can occur between the polymerases, and lead to the formation of recombined or
chimeric genomes13. Both of these things lead to the formation of quasispecies, where progeny
are created that are all of the PDV species, but they differ slightly in their genomic sequences13.
These differences, especially when they cause changes in the capsid protein, allow the virus to
change enough over time and make it harder for the immune system to recognize and destroy
them13.
Pathogenesis:
Since PDV is causing destruction of platelets, disorders such as thrombocytopenia
usually result in the host. This is a condition where abnormal amounts of bleeding happen
because the clotting system associated with the platelets is being removed4. This can cause
bleeding in the mouth and gums, bruising, nosebleeds, and a rash where red spots show up on the
body because of internal bleeding occurring4. The person though is typically asymptomatic, and
is unaware that they have been infected with the virus, until these unusual bleeding symptoms
start to occur. Internal bleeding or losing significant amounts of blood can lead to anemia or
weakness because of the iron loss and lack of oxygen transport4. However, because the virus only
has ways of attempting to avoid platelet immune functions when it is inside the cell, and not of
other immune cells when it is in the blood system, there are opportunities for the host innate and
adaptive immunity to combat the virus and keep it in check after some time. The time course and
outcome of this acute infection though is all dependent though on the types and amount of
genetic quasi-species the individual is exposed to, and how well the immune system can clear

them. If the immune system is successful though at combating the virus, and if the person is
given ways of dealing with lack of clotting and blood loss, such as blood or clotting factors, this
acute infection can be overcome over time. This outcome is the most likely, although if the
immune system has trouble overcoming it, then complications can arise from excessive bleeding,
especially if it occurs near major organs like the brain4. The virus typically though has ample
time to transfer to another host, even if the original host recovers from the thrombocytopenia and
is able to decrease the viral load to levels below that which can cause detectable symptoms.
Conclusion:
In conclusion, PDV is a positive sense RNA virus that attacks the platelet cells in
circulation in order to facilitate its life style. The goal of this virus is to cause acute bleeding
disorders in order to successfully transfer from host to host. It uses the aIIbb3integrinreceptor
to get into these megakaryocyte derived cells, utilizes their conserved protein synthesis
machinery to make more virion progeny, and then blows up the cell to find more potential
trophic cells to infect. The combination of its transcription and translation style, on top of useful
non-structural proteins like ISC allow it to overcome the platelet and try to gain the upper hand
in its host. Its destruction of platelets and speed of propagation allows it to cause a severe lack
of clotting and high viremia in the host within a small incubation period. While this virus and the
acute infection it causes can be overcome in most individuals, the virus is able to mutate quickly,
providing a challenge for the host immune system to detect and destroy it. Its function in other
mammals has not been discussed in this paper, but there is a potential for a mammalian wide
platelet tropism and infection, as platelets are a common feature amongst them1. Platelets,
particularly in other mammals, have not been thoroughly studied, so more information on the
potential of PDV infection still needs to be determined.

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