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Open AccessArticle

CPB1 of Aedes aegypti Interacts with DENV2 E Protein and Regulates Intracellular Viral Accumulation and Release from Midgut Cells

1
Virus-Host Interaction Research Group, Infectious Disease Laboratory, Jeffrey Cheah School of Medicine and Health Sciences, Monash University Malaysia, Jalan Lagoon Selatan, Bandar Sunway, 47500 Subang Jaya, Selangor, Malaysia
2
Department of Biotechnology and Neuroscience, Faculty of Life Science, Surya University, 15810 Tangerang, Banten, Indonesia
3
School of Biological Sciences, Universiti Sains Malaysia, 11800 USM, Pulau Pinang, Malaysia
*
Author to whom correspondence should be addressed.
Viruses 2014, 6(12), 5028-5046; https://doi.org/10.3390/v6125028
Received: 25 September 2014 / Revised: 24 November 2014 / Accepted: 11 December 2014 / Published: 16 December 2014
Aedes aegypti is a principal vector responsible for the transmission of dengue viruses (DENV). To date, vector control remains the key option for dengue disease management. To develop new vector control strategies, a more comprehensive understanding of the biological interactions between DENV and Ae. aegypti is required. In this study, a cDNA library derived from the midgut of female adult Ae. aegypti was used in yeast two-hybrid (Y2H) screenings against DENV2 envelope (E) protein. Among the many interacting proteins identified, carboxypeptidase B1 (CPB1) was selected, and its biological interaction with E protein in Ae. aegypti primary midgut cells was further validated. Our double immunofluorescent assay showed that CPB1-E interaction occurred in the endoplasmic reticulum (ER) of the Ae. aegypti primary midgut cells. Overexpression of CPB1 in mosquito cells resulted in intracellular DENV2 genomic RNA or virus particle accumulation, with a lower amount of virus release. Therefore, we postulated that in Ae. aegypti midgut cells, CPB1 binds to the E protein deposited on the ER intraluminal membranes and inhibits DENV2 RNA encapsulation, thus inhibiting budding from the ER, and may interfere with immature virus transportation to the trans-Golgi network. View Full-Text
Keywords: dengue virus; Aedes aegypti; carboxypeptidase; yeast two-hybrid dengue virus; Aedes aegypti; carboxypeptidase; yeast two-hybrid
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Tham, H.-W.; Balasubramaniam, V.R.M.T.; Tejo, B.A.; Ahmad, H.; Hassan, S.S. CPB1 of Aedes aegypti Interacts with DENV2 E Protein and Regulates Intracellular Viral Accumulation and Release from Midgut Cells. Viruses 2014, 6, 5028-5046.

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