Fig. 2: Nt-acetylation of Ser, Ala or Cys prevents them from recognition by ZER1 or ZYG11B.
From: CRL2ZER1/ZYG11B recognizes small N-terminal residues for degradation
a Schematic of the global protein stability (GPS) assay. PCMV, cytomegalovirus promoter, Ub ubiquitin, IRES internal ribosome entry site, GFP green fluorescent protein, RFP red fluorescent protein. b Stability analysis of GFLH-, SFLH- or AFLH-GFP upon ZER1 overexpression in HEK293T cells by GPS. The ratio of GFP/RFP was analyzed by flow cytometry. c Stability analysis of SFLH- or AFLH-fused GFP with full-length ZER1 overexpression in NAA10 knockdown cell lines. The ratio of GFP/RFP was analyzed by flow cytometry. d Stability analysis of N-terminal peptide derived from H2A with full-length ZER1 overexpression in NAA40 knockdown cell lines. The ratio of GFP/RFP was analyzed by flow cytometry. e Stability analysis of CFLH-fused GFP upon ZER1 or ZYG11B overexpression in HEK293T cells by GPS. The ratio of GFP/RFP was analyzed by flow cytometry. f Stability analysis of CFLH-fused GFP upon NAA10 knockdown or with simultaneous knockout of ZER1/ZYG11B in HEK293T cells. All FACS sequential gating images are provided in Supplementary Fig. 6.
