Zou Jiahui, Jiang Meijun, Xiao Rong, Sun Huimin, Liu Hailong, Peacock Thomas, Tu Shaoyu, Chen Tong, Guo Jinli, Zhao Yaxin, Barclay Wendy, Xie Shengsong, Zhou Hongbo
National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China.
Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education & Key Lab of Swine Genetics and Breeding of Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan, Hubei, People's Republic of China.
Nat Commun. 2025 Jan 15;16(1):670. doi: 10.1038/s41467-025-55903-0.
The Eurasian avian-like (EA) H1N1 swine influenza virus (SIV) possesses the capacity to instigate the next influenza pandemic, owing to its heightened affinity for the human-type α-2,6 sialic acid (SA) receptor. Nevertheless, the molecular mechanisms underlying the switch in receptor binding preferences of EA H1N1 SIV remain elusive. In this study, we conduct a comprehensive genome-wide CRISPR/Cas9 knockout screen utilizing EA H1N1 SIV in porcine kidney cells. Knocking out the enzyme gamma glutamyl carboxylase (GGCX) reduces virus replication in vitro and in vivo by inhibiting the carboxylation modification of viral haemagglutinin (HA) and the adhesion of progeny viruses, ultimately impeding the replication of EA H1N1 SIV. Furthermore, GGCX is revealed to be the determinant of the D225E substitution of EA H1N1 SIV, and GGCX-medicated carboxylation modification of HA 225E contributes to the receptor binding adaption of EA H1N1 SIV to the α-2,6 SA receptor. Taken together, our CRISPR screen has elucidated a novel function of GGCX in the support of EA H1N1 SIV adaption for binding to α-2,6 SA receptor. Consequently, GGCX emerges as a prospective antiviral target against the infection and transmission of EA H1N1 SIV.
欧亚禽源样（EA）H1N1猪流感病毒（SIV）具有引发下一次流感大流行的能力，这是由于其对人源α-2,6唾液酸（SA）受体具有更高的亲和力。然而，EA H1N1 SIV受体结合偏好转变背后的分子机制仍不清楚。在本研究中，我们利用EA H1N1 SIV在猪肾细胞中进行了全面的全基因组CRISPR/Cas9敲除筛选。敲除γ-谷氨酰羧化酶（GGCX）可通过抑制病毒血凝素（HA）的羧化修饰和子代病毒的黏附来降低病毒在体外和体内的复制，最终阻碍EA H1N1 SIV的复制。此外，GGCX被揭示为EA H1N1 SIV的D225E替代的决定因素，并且GGCX介导的HA 225E的羧化修饰有助于EA H1N1 SIV对α-2,6 SA受体的受体结合适应性。综上所述，我们的CRISPR筛选阐明了GGCX在支持EA H1N1 SIV适应结合α-2,6 SA受体方面的新功能。因此，GGCX成为针对EA H1N1 SIV感染和传播的潜在抗病毒靶点。
