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. 2018 Mar 26;115(15):E3529–E3538. doi: 10.1073/pnas.1721883115

Fig. 4.

Fig. 4.

Rep proteins interfere with the NIPP1–PP1 complex to antagonize KAP1-S824 dephosphorylation. (A) Schematic representation of NIPP1 mutants. The N-terminal FHA domain is shown in blue. FHAm denotes the FHA-binding mutant, which is unable to recruit hyperphosphorylated proteins. The central PP1-binding domain containing the consensus PP1-binding site RVTF is shown in yellow. RATA denotes the PP1-binding mutant. The C-terminal PP1 interaction and inhibitory domain are shown in green, and the RNA-binding region is shown in lime. (B) Cross-linked GFP-trap experiment performed in cells expressing Rep52 and each of the GFP-tagged NIPP1 constructs or a GFP control. (C) Immunoblot of p-KAP1-S824 in cells expressing NIPP1-WT, -RATA, -FHAm, or the empty vector (EV) control. (D) Quantification of p-KAP1-S824 levels in C. Values are reported as mean ± SEM, n = 4. (E) Immunoblot of p-KAP1-S824 in cells expressing Rep52 and NIPP1-WT, -RATA, -FHAm, or an empty vector (EV) control. (F) Quantification of p-KAP1-S824 levels in E. Values are reported as mean ± SEM, n = 5. (G) Cross-linked GFP trap performed in cells expressing NIPP1-WT-GFP and T7-tagged Rep52, Rep52K372A, or a Renilla control. Statistical significance was determined by unpaired t test, *P < 0.05.