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. 2023 Apr 1;299(5):104668. doi: 10.1016/j.jbc.2023.104668

Figure 1.

Figure 1

Screening of Hsp90α/β-interacting proteins among the 26 SARS-CoV-2 viral proteins.A, diagram of the SARS-CoV-2 genome and the names of all viral genes. Among the 16 nonstructural proteins (NSPs), NSP3 was dissected into its papain-like protease (PLpro) and ADP-ribose domains (ADRP) domains. The structural proteins are spike (S), membrane (M), envelope (E), and nucleoprotein (N). The accessory proteins are Orf3, Orf6, Orf7a, Orf7b, Orf8, and Orf9b. B–F, screening of Hsp90α/β interactors by coimmunoprecipitation (CoIP) assay. HEK293T cells were transfected with Hsp90β-flag (B and D) or Hsp90α-flag (C and D) vectors and myc-tagged SARS-CoV-2 proteins; total cellular proteins were extracted. CoIP assay was performed using an anti-flag (F3165) antibody and the CoIP protein complexes were screened by Western blot analysis using an anti-myc (71D10) antibody. HEK293T cells were transfected with pRK5-S (E) or E-mCherry (F) vectors and flag-tagged Hsp90α/β vectors; total cellular proteins were extracted. CoIP assay was performed using an anti-flag (F3165) antibody and the CoIP protein complexes were screened by Western blot analysis using an anti-S or anti-mCherry antibody. Proteins from lysates without pull-down confirmed the presence of the SARS-CoV-2 proteins as appropriate. Hsp90, heat shock protein 90; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2.