AUTHOR=Mao Haiying , Cao Lei , Xu Ting , Xia Xiaohan , Ren Peilei , Han Pengfei , Li Chengfei , Hui Xianfeng , Lin Xian , Huang Kun , Jin Meilin 

TITLE=YWHAG inhibits influenza a virus replication by suppressing the release of viral M2 protein

JOURNAL=Frontiers in Microbiology

VOLUME=Volume 13 - 2022

YEAR=2022

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2022.951009

DOI=10.3389/fmicb.2022.951009

ISSN=1664-302X

ABSTRACT=Influenza A virus (IAV) poses a serious threat to human life and property. The IAV matrix protein 2 (M2) plays an important role in viral budding. Increasing studies have proven the important roles of host factors in IAV replication. In this study, immunoprecipitation combined with mass spectrometry revealed that the host protein YWHAG, which YWHAG belongs to the 14-3-3 protein scaffold family, interacts with M2. Their interactions were further confirmed by co-immunoprecipitation, immunofluorescence and confocal microscopy of virus-infected HeLa cells. Moreover, we constructed YWHAG-KO and YWHAG-overexpressing cells and found that YWHAG knockdown significantly increased viral production, whereas its overexpression reduced the titer of virus progeny. Therefore, YWHAG is a negative regulatory factor during IAV infection. Further, YWHAG knockout or overexpression had no effect on the binding, entry, or viral RNA replication in the early stages of the virus life cycle. However, it impaired the release of virions at the plasma membrane as determined using transmission electron microscopy. We further found it suppressed the M2-mediated budding of the influenza virus. Importantly, the H158F mutation of YWHAG was found to affect its interaction with M2 and its budding. Collectively, our work demonstrates that YWHAG is a novel cellular regulator that targets and mediates the interaction and release of M2.