1. Interplay Between the Host Cell Membrane and dengue virus
.
Dengue virus (DENV) is an enveloped, positive sense single-stranded RNA virus
belonging to the Flavivirus genus. It is the causative agent of dengue fever, dengue
haemorrhagic and dengue shock syndrome, with 50 to 100 million people infected
annually. The number of cases of dengue fever and dengue haemorrhagic fever are also
on the rise.
DENV infection was found to induce the cytoplasmic membrane rearrangements, which
is vital for viral replication in host cells.1 In addition, DENV infection has been shown to
perturb cellular levels of HMG-CoA-reductase (HMGCR), the rate limiting enzyme in
cholesterol biosynthesis.2 However, it is unknown how modulation of cholesterol affects
the viral replication since both inhibition of HMGCR and addition of exogenous
cholesterol were found to reduce virus titer.2,3 Since cholesterol is a vital component of
the cell membrane, the perturbation of HMGCR could be associated with these
membrane rearrangements.
C6/36 cell line
infected with
DENV-2
This study aims to:
1. To identify the specific virus protein inducing replication vesicle formation in the host cell.
2. To reveal the key molecules – protein and lipid species – that induce and maintain
replication vesicle formation in the host cell.
3. To uncover the viral capacity to interfere with and manipulate basic cellular metabolism,
based on large-scale protein and lipid analysis by mass spectrometry.
UT-1 cells were
treated with
Compactin
Cellular Lipids,
Proteins analysis by
Mass Spectrometry
and cholesterol
concentration
assayed using Amplex
Red Cholesterol
Assay kit (Invitrogen)
Vs
Control
FixedforTEMat12,
24,36,48and72hr
control c6/36 cell line 24-hour-DENV-2Compactin treated UT-1 cells 12-hour – DENV-2
48-hour – DENV-2 72-hour – DENV-2
Cholesterol
0
50
100
150
Cholesterol(ug/ml)
Cholesterol Ester
0
20
40
60
80
CholesterolEsters(ug/ml)
Department Of Biotechnology Indian Academy Centre for Research & P.G. Studies Bangalore
Email Id: rashmirekha978@gmail.com
Rashmirekha Mohapatra1, Sarshad Ibnu Sayed, Selvam Arjunan.
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0
Time, min
0.0
2.0e4
4.0e4
6.0e4
8.0e4
1.0e5
1.2e5
1.4e5
Intensity,cps
Cholesterol
Cholesterol Esters
Control
Drug Treated
• DENV-2 infection increased the level of cholesterol in a time-dependent manner.
• DENV-2 infection leads to membrane arrangements, autophagy interaction and lipid
droplet
1. S. Welsch et al., Cell Host Microbe 5, 365 (2009).
2. C. Rothwell et al., Virology 389, 8 (2009).
3. C. J. Lee et al., J Virol 82, 6470 (2008).
4. L. M. Pfeffer et al., Proc Natl Acad Sci U S A 82, 2417 (1985).
5. J. Y. Luu et al., Hum Pathol 20, 617 (1989).
1. To verify the key molecule (proteins/lipids) in the induced membrane formation
after DENV-2 infection in c6/36 cell lines.
2.To elucidate change in cholesterol localization induced by DENV-2.
I would like to express my gratitude to the UGC for funding and National Institute of virology for providing
technical assistance on laboratory and NIMHANS for electron Microscopy. I would also like to thank the
members of department of biotechnology of Indaian Academy college for their support.