2016 Fiscal Year Research-status Report
Studies on influenza A virus M2-host interactome and its role in virus replication
| Project/Area Number |
16K08014
|
|---|
| Research Institution | Hokkaido University |
|---|
Principal Investigator |
マンズール ラシッド 北海道大学, 人獣共通感染症リサーチセンター, 特任助教 (90566150)
|
|---|
| Project Period (FY) |
2016-04-01 – 2019-03-31
|
| Keywords | Influenza A virus / M2 protein |
|---|
| Outline of Annual Research Achievements |
The viruses upon infection hijack the cell machinery and re-route or modulate it to complete their life cycle. Therefore, viruses or their encoded proteins must interact with host proteins for successful completion of their life cycle. Despite the important role of influenza A virus (IAV) M2 protein in virus life cycle such as, vRNP release, virus morphogenesis and budding, very little information on M2-host interactome is available. In this study, we plan to investigate the role of identified M2-interacting host proteins in virus replication.
During this fiscal year, siRNA based screening was conducted using three siRNAs per target gene. Initially screening conditions such as optimal virus dose, siRNA dose, and suitable cell line (293T, A549 etc) were optimized. The host factors were shortlisted by determining the gene knockdown effect on virus infectivity. The effect of gene knockdown on virus replication was considered positive or negative when at least 2/3 siRNAs produced similar effect. So far, knockdown of five genes caused more than thirty percent reduction in infectivity and three genes caused increase in virus infectivity.
|
| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Further investigations are being conducted to study the role of shortlisted host proteins in virus replication. Members of solute carrier family proteins i.e. SLC25A5, SLC1A5 are among the shortlisted candidate proteins. At present, there putative role in virus replication is being investigated.
To study the effect of gene knockdown on virus budding, an influenza virus has been generated by reverse genetics. Moreover, a stable cell line expressing M2 protein is being developed to study the effect of gene knockdown on M2 cytotoxicity.
|
| Strategy for Future Research Activity |
1. To continue to investigate the role of shortlisted host proteins in influenza virus replication with relevance to M2 protein function.
2. To optimize 2D electrophoresis for studying the host plasma membrane proteome in M2 expressing cells.
|
| Causes of Carryover |
Since the preparation of rg-virus and stable cell line required for screening of host factors in virus budding were delayed; therefore, the consumables and reagents required could not be purchased during this fiscal year.
|
| Expenditure Plan for Carryover Budget |
The unused amount will be used to continue the study in the next fiscal year. The money will be used for buying the consumables necessary for execution of study such as siRNAs, antibodies, media, plastic ware etc. Additionally, the amount will be used for presenting the research results.
|
-
[Journal Article] Genetic Predisposition To Acquire a Polybasic Cleavage Site for Highly Pathogenic Avian Influenza Virus Hemagglutinin2017
Author(s)
Naganori Nao, Junya Yamagishi, Hiroko Miyamoto, Manabu Igarashi, Rashid Manzoor, Aiko Ohnuma, Yoshimi Tsuda, Wakako Furuyama, Asako Shigeno, Masahiro Kajihara, Noriko Kishida, Reiko Yoshida, Ayato Takada
-
Journal Title
mBio
Volume: 8
Pages: 1-15
DOI
Peer Reviewed / Open Access
-
-
