tetano
Editor, Senior Moderator
Biochem Biophys Res Commun
. 2025 Apr 29:767:151915.
doi: 10.1016/j.bbrc.2025.151915. Online ahead of print. High-resolution structure reveals enhanced 14-3-3 binding by a mutant SARS-CoV-2 nucleoprotein variant with improved replicative fitness
Kristina V Perfilova[SUP] 1 [/SUP], Ilya O Matyuta[SUP] 2 [/SUP], Mikhail E Minyaev[SUP] 3 [/SUP], Konstantin M Boyko[SUP] 1 [/SUP], Richard B Cooley[SUP] 4 [/SUP], Nikolai N Sluchanko[SUP] 5 [/SUP]
Affiliations
Replication of many viruses depends on phosphorylation of viral proteins by host protein kinases and subsequent recruitment of host protein partners. The nucleoprotein (N) of SARS-CoV-2 is heavily phosphorylated and recruits human phosphopeptide-binding 14-3-3 proteins early in infection, which is reversed prior to nucleocapsid assembly in new virions. Among the multiple phosphosites of N, which are particularly dense in the serine/arginine-rich interdomain region, phospho-Thr205 is highly relevant for 14-3-3 recruitment by SARS-CoV-2 N. The context of this site is mutated in most SARS-CoV-2 variants of concern. Among mutations that increase infectious virus titers, the S202R mutation (B.1.526 Iota) causes a striking replication boost (∼166-fold), although its molecular consequences have remained unclear. Here, we show that the S202R-mutated N phosphopeptide exhibits a 5-fold higher affinity for human 14-3-3ζ than the Wuhan variant and we rationalize this effect by solving a high-resolution crystal structure of the complex. The structure revealed an enhanced 14-3-3/N interface contributed by the Arg202 side chain that, in contrast to Ser202, formed multiple stabilizing contacts with 14-3-3, including water-mediated H-bonds and guanidinium pi-pi stacking. These findings provide a compelling link between the replicative fitness of SARS-CoV-2 and the N protein's affinity for host 14-3-3 proteins.
Keywords: Coronavirus; Crystal structure; Phosphorylation; Variant of concern; Viral replication.
. 2025 Apr 29:767:151915.
doi: 10.1016/j.bbrc.2025.151915. Online ahead of print. High-resolution structure reveals enhanced 14-3-3 binding by a mutant SARS-CoV-2 nucleoprotein variant with improved replicative fitness
Kristina V Perfilova[SUP] 1 [/SUP], Ilya O Matyuta[SUP] 2 [/SUP], Mikhail E Minyaev[SUP] 3 [/SUP], Konstantin M Boyko[SUP] 1 [/SUP], Richard B Cooley[SUP] 4 [/SUP], Nikolai N Sluchanko[SUP] 5 [/SUP]
Affiliations
- PMID: 40318379
- DOI: 10.1016/j.bbrc.2025.151915
Replication of many viruses depends on phosphorylation of viral proteins by host protein kinases and subsequent recruitment of host protein partners. The nucleoprotein (N) of SARS-CoV-2 is heavily phosphorylated and recruits human phosphopeptide-binding 14-3-3 proteins early in infection, which is reversed prior to nucleocapsid assembly in new virions. Among the multiple phosphosites of N, which are particularly dense in the serine/arginine-rich interdomain region, phospho-Thr205 is highly relevant for 14-3-3 recruitment by SARS-CoV-2 N. The context of this site is mutated in most SARS-CoV-2 variants of concern. Among mutations that increase infectious virus titers, the S202R mutation (B.1.526 Iota) causes a striking replication boost (∼166-fold), although its molecular consequences have remained unclear. Here, we show that the S202R-mutated N phosphopeptide exhibits a 5-fold higher affinity for human 14-3-3ζ than the Wuhan variant and we rationalize this effect by solving a high-resolution crystal structure of the complex. The structure revealed an enhanced 14-3-3/N interface contributed by the Arg202 side chain that, in contrast to Ser202, formed multiple stabilizing contacts with 14-3-3, including water-mediated H-bonds and guanidinium pi-pi stacking. These findings provide a compelling link between the replicative fitness of SARS-CoV-2 and the N protein's affinity for host 14-3-3 proteins.
Keywords: Coronavirus; Crystal structure; Phosphorylation; Variant of concern; Viral replication.