Fig. 5: Lysine 51 of PSAT1 is an essential site for the interaction with USP14. | Communications Biology

Fig. 5: Lysine 51 of PSAT1 is an essential site for the interaction with USP14.

From: Interplay between acetylation and ubiquitination controls PSAT1 protein stability in lung adenocarcinoma

Fig. 5

a The co-modified sites with ubiquitination and acetylation of PSAT1 predicted by PhosphoSitePlus. b The expression effects of PSAT1 plasmids containing indicated mutations (K51R, K311R, K323R, K333R and K363R) were tested. c–g H1299 cells were co-transfected with His-USP14 and Flag-PSAT1K51R, Flag-PSAT1K311R, Flag-PSAT1K323R, Flag-PSAT1 K333R or Flag-PSAT1K363R plasmids. Co-IP was used to detect the total ubiquitination (left panel) and K48-linked ubiquitination (right panel) levels of PSAT1. h H1299 cells were experimented with CHX (100 μg/mL) for various time points and were transfected with Flag-PSAT1WT or Flag-PSAT1K51R. The Flag-PSAT1WT or Flag-PSAT1K51R protein expressions were detected by Western blot. The relative protein expression compared with that of Tubulin was quantified. Data represented as the average of three independent experiments (mean ± SD), *P < 0.05. i H1299 cells were co-transfected with His-USP14, Flag-PSAT1WT or Flag-PSAT1K51R. The immunoprecipitations were blotted with anti-His or anti-Flag antibodies. The relative His-USP14 expression compared with that of Flag was quantified. Data represented as the average of three independent experiments (mean ± SD), ***P < 0.001.

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