Genetic Engineering Research Laboratory, Beijing Institute of Biotechnology, Beijing, China.
Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun, China.
J Virol. 2024 Apr 16;98(4):e0156523. doi: 10.1128/jvi.01565-23. Epub 2024 Mar 6.
The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has posed a worldwide threat in the past 3 years. Although it has been widely and intensively investigated, the mechanism underlying the coronavirus-host interaction requires further elucidation, which may contribute to the development of new antiviral strategies. Here, we demonstrated that the host cAMP-responsive element-binding protein (CREB1) interacts with the non-structural protein 13 (nsp13) of SARS-CoV-2, a conserved helicase for coronavirus replication, both in cells and in lung tissues subjected to SARS-CoV-2 infection. The ATPase and helicase activity of viral nsp13 were shown to be potentiated by CREB1 association, as well as by Protein kinase A (PKA)-mediated CREB1 activation. SARS-CoV-2 replication is significantly suppressed by PKA Cα, cAMP-activated protein kinase catalytic subunit alpha (PRKACA), and CREB1 knockdown or inhibition. Consistently, the CREB1 inhibitor 666-15 has shown significant antiviral effects against both the WIV04 strain and the Omicron strain of the SARS-CoV-2. Our findings indicate that the PKA-CREB1 signaling axis may serve as a novel therapeutic target against coronavirus infection.
In this study, we provide solid evidence that host transcription factor cAMP-responsive element-binding protein (CREB1) interacts directly with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) helicase non-structural protein 13 (nsp13) and potentiate its ATPase and helicase activity. And by live SARS-CoV-2 virus infection, the inhibition of CREB1 dramatically impairs SARS-CoV-2 replication . Notably, the IC50 of CREB1 inhibitor 666-15 is comparable to that of remdesivir. These results may extend to all highly pathogenic coronaviruses due to the conserved nsp13 sequences in the virus.
由严重急性呼吸系统综合症冠状病毒 2（SARS-CoV-2）引起的 COVID-19 大流行在过去 3 年中构成了全球性威胁。尽管已经进行了广泛而深入的研究，但冠状病毒与宿主相互作用的机制仍需要进一步阐明，这可能有助于开发新的抗病毒策略。在这里，我们证明宿主 cAMP 反应元件结合蛋白（CREB1）与 SARS-CoV-2 的非结构蛋白 13（nsp13）相互作用，nsp13 是冠状病毒复制的一种保守解旋酶，无论是在细胞中还是在 SARS-CoV-2 感染的肺组织中。实验表明，病毒 nsp13 的 ATP 酶和解旋酶活性可通过 CREB1 结合以及蛋白激酶 A（PKA）介导的 CREB1 激活得到增强。PKA Cα、cAMP 激活蛋白激酶催化亚基α（PRKACA）和 CREB1 敲低或抑制显著抑制 SARS-CoV-2 复制。一致地，CREB1 抑制剂 666-15 对 WIV04 株和 SARS-CoV-2 的奥密克戎株均显示出显著的抗病毒作用。我们的研究结果表明，PKA-CREB1 信号通路可能是针对冠状病毒感染的新型治疗靶标。
在这项研究中，我们提供了确凿的证据表明，宿主转录因子 cAMP 反应元件结合蛋白（CREB1）与严重急性呼吸系统综合症冠状病毒 2（SARS-CoV-2）解旋酶非结构蛋白 13（nsp13）直接相互作用，并增强其 ATP 酶和解旋酶活性。通过活 SARS-CoV-2 病毒感染，CREB1 的抑制显著损害了 SARS-CoV-2 的复制。值得注意的是，CREB1 抑制剂 666-15 的 IC50 与瑞德西韦相当。由于病毒中 nsp13 序列的保守性，这些结果可能扩展到所有高致病性冠状病毒。
