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Screening of host proteins interacting with Kunjin, Langat, Zika replication complex
Örebro University, School of Medical Sciences. (Molecular Flavirus group)
Örebro University, School of Medical Sciences.
Department ofClinical Microbiology, Sahlgrenska University Hospital, Gothenburg, Sweden; NanoxisConsulting AB, Gothenburg, Sweden.
Department ofClinical Microbiology, Sahlgrenska University Hospital, Gothenburg, Sweden; NanoxisConsulting AB, Gothenburg, Sweden.
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2019 (English)In: Positive-Strand Rna Viuses, 2019Conference paper, Poster (with or without abstract) (Refereed)
Abstract [en]

When Flaviviruses infect host cells, they can induce invagination of endoplasmic reticulum (ER) membrane to form vesicle-like compartments. These unique structures are hypothetical to facilitate the viral replication by reducing diffusion of virus replication machinery and viral RNA, providing a scaffold to anchor the replication complex, and protecting viral RNA from host cell intrinsic surveillance. 

The rearrangements of ER membrane to form these replication compartments (RCs) require modifications in its lipid constituents or binding of proteins to the membrane. Flaviviruses, indeed, use their proteins to generate RCs. It has been implicated that both KUNV and DENV viral NS1, NS2A, NS4A, NS4B proteins could induce membrane remodelings. However, it is recondite whether host proteins can also participate in the formation and maintenance of RCs.

In this project, we aimed to identify and characterize of host proteins inducing RC generation during Flavivirus infections. We used A549 as a cell model, and mosquito-borne Zika and Kunjin virus, and tick-borne Langat virus as virus models. After virus infections, ER membranes were harvested using ultracentrifuge with a sucrose gradient. Proteins from these ERs were identified using mass spectrometry. We compared the differences between the ER proteomes of infected cells and non-infected cells to identify host candidate proteins that can cause the RC formation.  We are attempting to enrich the RC-containing fractions and identifying proteins here, which narrows the list of true candidate proteins. The candidate proteins then will be characterized by using molecular techniques such as gene knock down, overexpression, and microscopy techniques.

Place, publisher, year, edition, pages
2019.
Keywords [en]
Replication complex, Kunjin, Langat, Zika
National Category
Medical and Health Sciences Infectious Medicine
Research subject
Molecular Cellbiology; Biomedicine; Infectious Diseases
Identifiers
URN: urn:nbn:se:oru:diva-76400OAI: oai:DiVA.org:oru-76400DiVA, id: diva2:1351273
Conference
Positive-Strand RNA Viuses, KILLARNEY, Co.Kerry, Ireland, June 9-13,2019
Funder
Knowledge Foundation, HÖG15 20150201Available from: 2019-09-13 Created: 2019-09-13 Last updated: 2019-09-13Bibliographically approved

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Screening of host proteins interacting with Kunjin, Langat, Zikareplication complex(1237 kB)12 downloads
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Tran, Pham Tue HungAsghar, NaveedJohansson, MagnusMelik, Wessam

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