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. Author manuscript; available in PMC: 2021 Dec 3.
Published in final edited form as: Cell Rep. 2021 Oct 19;37(3):109840. doi: 10.1016/j.celrep.2021.109840

Figure 2. Dfm1 is intolerable to mutations in Loop 1 (L1) and transmembrane domain 2 (TM2).

Figure 2.

(A) Depiction of fusion protein, SUS-GFP. The transmembrane Hmg1 domain has a luminal Myc epitope and cytosolic 3×HA epitopes fused to the catalytic Hrd1 RING.

(B) Mutagenized DFM1 was transformed into dfm1Δ hrd1Δ cells expressing SUS-GFP and scored for stabilization of SUS-GFP or high colony fluorescence by visualization.

(C) Depiction of Dfm1 mutants (indicated in red for L1 and green for TM2) that were selected from the random mutagenesis screen and validated for expression, localization to ER, and Cdc48 recruitment function.

(D) Homology model of Dfm1. Positions of L1 and TM2 mutants are indicated in red and green, respectively.