tetano
Editor, Senior Moderator
Commun Biol
. 2022 Sep 7;5(1):925.
doi: 10.1038/s42003-022-03892-x.
Multiscale characterization reveals oligomerization dependent phase separation of primer-independent RNA polymerase nsp8 from SARS-CoV-2
Jinxin Xu[SUP] #[/SUP][SUP] 1 2 [/SUP], Xin Jiang[SUP] #[/SUP][SUP] 3 [/SUP], Yulong Zhang[SUP] 1 2 [/SUP], Yan Dong[SUP] 1 [/SUP], Changli Ma[SUP] 4 [/SUP], Hanqiu Jiang[SUP] 4 [/SUP], Taisen Zuo[SUP] 4 [/SUP], Rui Chen[SUP] 1 5 [/SUP], Yubin Ke[SUP] 4 [/SUP], He Cheng[SUP] 4 [/SUP], Howard Wang[SUP] 6 [/SUP], Jinsong Liu[SUP] 7 8 9 10 11 [/SUP]
Affiliations
Abstract
RNA replication and transcription machinery is an important drug target for fighting against coronavirus. Non-structure protein nsp8 was proposed harboring primase activity. However, the RNA primer synthesis mechanism of nsp8 is still largely unknown. Here, we purified dimer and tetramer forms of SARS-CoV-2 nsp8. Combined with dynamic light scattering, small-angle neutron scattering and thermo-stability analysis, we found that both dimer and tetramer become loosened and destabilized with decreasing salt concentration, and the dimer form is more stable than the tetramer form. Further investigation showed that nsp8 dimer and tetramer can undergo phase separation but exhibit different phase separation behaviors. Nsp8 dimer can form liquid-like droplets in the buffer with a low concentration of NaCl; phase separation of nsp8 tetramer depends on the assistance of RNA. Our findings on different phase separation behaviors of nsp8 dimer and tetramer may provide insight into the functional studies of nsp8 in coronavirus.
. 2022 Sep 7;5(1):925.
doi: 10.1038/s42003-022-03892-x.
Multiscale characterization reveals oligomerization dependent phase separation of primer-independent RNA polymerase nsp8 from SARS-CoV-2
Jinxin Xu[SUP] #[/SUP][SUP] 1 2 [/SUP], Xin Jiang[SUP] #[/SUP][SUP] 3 [/SUP], Yulong Zhang[SUP] 1 2 [/SUP], Yan Dong[SUP] 1 [/SUP], Changli Ma[SUP] 4 [/SUP], Hanqiu Jiang[SUP] 4 [/SUP], Taisen Zuo[SUP] 4 [/SUP], Rui Chen[SUP] 1 5 [/SUP], Yubin Ke[SUP] 4 [/SUP], He Cheng[SUP] 4 [/SUP], Howard Wang[SUP] 6 [/SUP], Jinsong Liu[SUP] 7 8 9 10 11 [/SUP]
Affiliations
- PMID: 36071105
- DOI: 10.1038/s42003-022-03892-x
Abstract
RNA replication and transcription machinery is an important drug target for fighting against coronavirus. Non-structure protein nsp8 was proposed harboring primase activity. However, the RNA primer synthesis mechanism of nsp8 is still largely unknown. Here, we purified dimer and tetramer forms of SARS-CoV-2 nsp8. Combined with dynamic light scattering, small-angle neutron scattering and thermo-stability analysis, we found that both dimer and tetramer become loosened and destabilized with decreasing salt concentration, and the dimer form is more stable than the tetramer form. Further investigation showed that nsp8 dimer and tetramer can undergo phase separation but exhibit different phase separation behaviors. Nsp8 dimer can form liquid-like droplets in the buffer with a low concentration of NaCl; phase separation of nsp8 tetramer depends on the assistance of RNA. Our findings on different phase separation behaviors of nsp8 dimer and tetramer may provide insight into the functional studies of nsp8 in coronavirus.