Figure 2: Npr1 controls Mep2 activity via S457 phosphorylation. | Nature Communications

Figure 2: Npr1 controls Mep2 activity via S457 phosphorylation.

From: The TORC1 effector kinase Npr1 fine tunes the inherent activity of the Mep2 ammonium transport protein

Figure 2

(a,b) Triple-mepΔ (31019b) and triple-mepΔ npr1-1 (31052c) cells were transformed with YCpMep2N4Q, YCpMep2N4Q,S457A,T459A,S460A, YCpMep2N4Q,S457A,T459A, YCpMep2N4Q,S457A,S460A, YCpMep2N4Q,T459A,S460A, YCpMep2N4Q,S457A, YCpMep2N4Q,S460A, YCpMep2N4Q,S457D, or with YCpMep2N4Q,S460D. (a) Immunodetection of total Mep2N4Q and S457-phosphorylated Mep2N4Q (pS457-Mep2) from membrane-enriched extracts of proline-grown cells. Pma1 was detected as a loading control. (b) Growth tests on solid medium containing, as the sole nitrogen source, 1 mM ammonium (Am) or 0.1% glutamate (Glt, positive growth control). Cells were incubated at 29 °C for 4 days. (c) Immunodetection of pS457-Mep2 using the same membrane as in Fig. 1a. (d) Immunodetection of Mep2N4Q and pS457-Mep2 from membrane-enriched extracts of proline-grown triple-mepΔ (31019b) cells transformed with YCpMep2N4Q, YCpMep2N4Q,S457A, YCpMep2N4Q,S460A, YCpMep2N4Q,S457D, or with YCpMep2N4Q,S460D. (e) Immunodetection of pS457-Mep2 using the same membrane as in Fig. 1b.

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