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Group 1 Presentation
1. STACY JACOBS (GROUP 1)
Aedes aegypti’s Regulation of Genes Impact
the Spread of Dengue Infections Through
Alterations to Dengue Virus
Replication/TransmissionA
2. Overview
Group 3 Group 2 Group 1 Group 4
• Aedes aegypti
• Life Cycle
-Adult
-Aquatic
• Morphology
• wMelPop
• Histology
• Transmission of
Dengue
-Mouth to
midgut
-Midgut to
hemocoel
-Hemocoel to
all organs
-Salivary
Glands
• Transcription
• Translation
• Dengue Virus
Replication
• ADE
• Genes
• Toll Pathway
• Habitat
-Temperature
-Location
• Keystone
Species
• Trophic Levels
• Prevention
Methods
6. Virus4,J
Composed mainly of
RNA/DNA
Enclosed in a protein shell
Hijack cellular machinery
of host cells
Inject genetic material into
host cells
Structural rearrangement
of the viral protein coat
7. Dengue Virus4,D
Enveloped positive strand RNA virus
Structural Proteins
-Capsid (C) Protein
-Membrane (M) Protein
-Envelope (E) Protein
Viral envelope surrounds the nucleocapsid
E and M proteins attach to viral envelope
8. Dengue Virus Serotypes2,4,I
Four Serotypes:
DENV 1, 2, 3, & 4
Genetic variation
within serotypes due
to difference in
antigens
9. Dengue Virus Serotypes2,4
Provides specific lifetime immunity and
short-term cross-immunity
Some genetic variants within each
serotype may be more virulent or have
greater epidemic potential
Bangkok, Thailand: 1994-2006
-DENV-1: more common
-DENV-2: more virulent
10. Dengue E Protein Dimer4
Domain I = red
Domain II = yellow
Domain III = blue
Acidic pH -> fusion peptides (in green) are exposed
to target membrane
Domain III folds toward the fusion peptides
Forcing the target membrane and viral membrane to
bend toward each other and fuse
12. Dengue Virus Transmission4,E
Clatherin-mediated
endocytosis
Nucleocapsid is
uncoated
RNA is translated and
folded
New RNA is packaged
into a nucleocapsid
13. Dengue Virus Transmission4,E
• Nucleocapsid enters
the ER -> translates
proteins; budding
occurs
• Nucleocapsid enters
the Golgi -> furin
cleavage
• Immature virus
matures and exits the
cell via exocytosis
16. Antibody-Dependent Enhancement3,G
One serotype infects an
individual; later
another serotype
infects the same
individual
Results in higher
viremia
Secondary infections
tend to cause more
severe symptoms
17. Aedes aegypti’s Genome Map5
1,376 Mb
Four Quantitative
Trait Locis
(QTLs) related to
the transmission
of dengue
encompassed 11%
of chromosome 2
18. Regulation of Genes1
Differentially Up-Regulated Genes (DURGs):
when a cell is deficient, more receptor protein is
synthesized
Differentially Down-Regulated Genes
(DDRGs): when a cell is overstimulated, the
expression of the receptor protein is decreased
19. Results – DDRGs1
AAEL011045 gene Pupal
Cuticle (PC) Protein
AAEL003012 gene Matrix
Metalloprotease (MMP) for zinc
Overexpression causes
flaviviruses to be inhibited one
million fold in mosquitoes
20. Results – DDRGs1
PC Protein binds E protein on a virus -> inhibits
infection in mosquitoes and mice
MMP inhibits infection in mosquitoes, but not in
mice
21. Results – DURG1
AAEL014440 gene Juvenile Hormone
Inducible Protein
- up-regulated at all time points for all flaviviruses
- regulates many other genes
AAEL003685 gene Core Histone H3 Protein
- 4 fold up-regulated at all time points for all
flaviviruses
23. Toll Pathway6
Uses a large number of DDRGs and DURGs that
function in immune response
-34.5% in midgut
-27.5% in carcass
Myeloid Differentiation Primary Response gene 88
(MYD88) Cytoplasmic Adaptor Protein
Cytoplasmic Adaptor Protein binds to a receptor ->
activates Toll Pathway
24. Toll Pathway6
When MYD88 is silenced,
Toll Pathway is repressed.
Therefore, dengue has
higher rates of infection.
When the Cactus gene is
activated, the Toll Pathway
is stopped.
25. Concluding Remarks
Dengue Infection can be controlled by:
- Alteration to Dengue Replication and
Transmission -> Change in pH, Antibody
Dependent Enhancement (ADE)
- Overexpression of DDRGs in A. aegypti
- MYD88 expression to activate Toll
Pathway
- Inhibition of Cactus does not stop Toll
Pathway
26. Works Cited - Literature
1. Colpitts, T., Cox, J., Vanlandingham, D., Feitosa, F., Cheng, G., Kurscheid, S., Wang, P.,
Krishnan, M., Higgs, S. and Firkrig, E. 2011. Alterations in the Aedes aegypti
transcriptome during infection with West Nile, dengue and yellow fever
viruses. PLoS Pathogens 7, e1002189.
2. Fried, J., Gibbons, R., Kalayanarooj, S., Thomas, S., Srikiatkhachorn, A., Yoon, I-K.,
Jarman, R., Green, S., Rothman, A. and Cummings, D. 2010. Serotype-specific
differences in the risk of dengue hemorrhagic fever: An analysis of data
collected in Bangkok, Thailand from 1994 to 2006. PLoS Neglected Tropical
Diseases 4, e617.
3. Grandi, G. 2007. In vitro transcription and translation protocols. Totoway, NJ.
Humana Press.
4. Rodenhuis-Zybert, I., Wilschut, J. and Smit, J. 2010. Dengue virus life cycle: viral and
host factors modulating infectivity. Cellular and Molecular Life Sciences 67,
2773-2786.
5. Timoshevskiy, V., Severson, D., deBruyn, B., Black, W., Sharakhov, I. and Sharakhov,
M. 2013. An integrated linkage, chromosome, and genome map for the yellow
fever mosquito Aedes aegypti. PLoS Neglected Tropical Diseases 7, e2052.
6. Zhiyong, X., Ramirez, J. and Dimopoulos G. 2008. The Aedes aegypti Toll pathway
controls dengue virus infection. PLoS Pathogens 4, e1000098.
27. Works Cited - Images
A. http://www.nowpublic.com/health/aedes-aegypti-0
B. http://hubpages.com/hub/protein-production-a-step-by-step-illustrated-
guide
C. http://denydendhi.blogspot.com/2011/03/replikasi-dna.html
D. http://www.nature.com/scitable/topicpage/dengue-viruses-22400925
E.http://pic1.gophoto.us/key/dengue%20virus%20life%20cycle%20ppt
F.http://www.pnas.org/content/109/1/E23/F6.expansion.html
G.http://www.niaid.nih.gov/labsandresources/labs/aboutlabs/lvd/viralpathog
enesissection/Pages/default.aspx
H.http://pioneerbiology.wordpress.com/2010/10/24/transcriptiontransation/
I.http://www.biology.arizona.edu/immunology/tutorials/antibody/structure.ht
ml
J. http://oceanworld.tamu.edu/resources/oceanography-
book/microbialweb.htm
Editor's Notes
vesicles containing proteins with receptor sites specific to the molecules being internalized.
Furin is a protein that cleaves precursor proteins at their paired basic amino acid processing sites
Fc Receptors are found on macrophages, monocytes, dendritic cells
However, domain I and II are targeted by human antibodies.