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. 2021 Oct 6;40(22):e107958. doi: 10.15252/embj.2021107958

Figure 6. Ice2 opposes Pah1 by inhibiting the Nem1‐Spo7 complex.

Figure 6

  1. Schematic of Pah1 phospho‐regulation. Phosphorylated Pah1 is cytosolic and inactive. Interaction of Pah1 and the ER‐localized Nem1‐Spo7 complex results in Pah1 dephosphorylation and activation, promoting conversion of PA into DAG.
  2. Western blot of HA from WT, Δice2, Δnem1, and Δnem1 Δice2 cells expressing endogenously tagged Pah1‐HA (SSY2592, 2593, 2594, 2718). Blots of SDS‐PAGE and Phos‐tag PAGE gels are shown.
  3. Schematic of Pah1 dephosphorylation assay with phosphorylated Pah1 from ∆nem1 mutants and microsomes from different strains.
  4. Western blot of HA from Pah1 dephosphorylation reactions that contained phosphorylated Pah1‐HA from ∆nem1 mutants (SSY3065) and microsomes from cells of the indicated genotypes (SSY3053, 3074, 3075, 3095). Phosphorylated Pah1 and dephosphorylated Pah1 resulting from treatment with recombinant alkaline phosphatase (PPase) are shown for reference.
  5. Western blot of HA, Sec61, and Pgk1 from cell lysates and microsomes prepared from WT and ∆ice2 cells expressing Nem1‐HA (SSY3140, 3141). Nem1 is undetectable in cell lysates due to its low abundance.
  6. Western blot of HA from Pah1 phosphorylation reaction that contained hypophosphorylated Pah1‐HA from ∆ice2 cells (SSY3096) and microsomes from ∆nem1 cells (SSY3075). Phosphorylated Pah1 from ∆nem1 cells (SSY3065) and hypophosphorylated Pah1 from ∆ice2 cells are shown for reference.

Source data are available online for this figure.