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. Author manuscript; available in PMC: 2023 Jul 10.
Published in final edited form as: Sci Signal. 2023 Jan 10;16(767):eabm5518. doi: 10.1126/scisignal.abm5518

Figure 1. A system coupling MAPK D-site interactions to yeast cell growth.

Figure 1.

(A) Scheme showing the impact of replacing yeast MAPK pathway components with human homologs on cell growth. (B) Domain structure of MKK6 and D-site variants. MKK6D7 replaces the native p38-selective D-site with that from MKK7, which binds only to JNK. (C) Growth assay for a hog1Δ pbs2Δ strain co-expressing p38α and the indicated MKK6 variants. Cells were grown in liquid culture, derepressed in raffinose media, and then spotted in 5-fold serial dilutions on solid media containing either galactose (to induce p38 expression) or glucose. Representative of 2 independent experiments. (D) Growth assay of a hog1Δ pbs2Δ strain co-expressing JNK1 and MKK6 variants conducted as in (C). Representative of 2 independent experiments. (E) Scheme showing potential mechanisms for growth rescue resulting from the expression of GST-D-site fusion proteins. (F) Effect of expressing cognate (MKK6, MEF2A) or non-cognate (MKK7) D-sites fused to GST on growth arrest mediated by p38α-MKK6 co-expression in a hog1Δ pbs2Δ strain. Cells were grown and plated as in (C). Representative of 2 independent experiments. (G) Effect of expressing cognate (MKK7, NFAT4) or non-cognate (MKK6) D-sites fused to GST on growth arrest mediated by JNK1-MKK6D7 co-expression in a hog1Δ pbs2Δ strain. Representative of 2 independent experiments.