Fig. 2: SaciUrm1 conjugates to yeast Ahp1 in an urmylation-like fashion. | Communications Biology

Fig. 2: SaciUrm1 conjugates to yeast Ahp1 in an urmylation-like fashion.

From: Evolutionary conservation of ubiquitin-like protein urmylation as revealed by URM1 gene shuffle from archaea to yeast

Fig. 2: SaciUrm1 conjugates to yeast Ahp1 in an urmylation-like fashion.

A, B EMSA under reducing and conjugating conditions. Protein extracts from urm1∆, urm1∆AHP1-c-MYC or urm1∆ahp1∆ strains expressing HA-SaciURM1 (A) or HA-ScURM1 (B) were subjected to anti-HA Western blots (top panels) to detect free HA-SaciUrm1 (~20 kDa) or HA-ScUrm1 (~23 kDa) and urmylated Ahp1 (~40 kDa) or Ahp1-c-Myc (~50 kDa) conjugates. Unconjugated Ahp1 (~20 kDa) or Ahp1-c-Myc (~25 kDa) is monitored in anti-Ahp1 Western blots (middle panels). C Urmylation analysis of Ahp1 mutants by EMSA. Protein extracts from urm1∆ahp1∆ strains co-expressing HA-SaciURM1 and indicated AHP1 substitutions or carrying empty vector (ev) were subjected to anti-HA Western blots (top panel). Free forms of HA-SaciUrm1 (~20 kDa) and urmylated Ahp1 (~40 kDa) conjugates are marked. Free Ahp1 (~20 kDa) is detected by an anti-Ahp1 blot (middle panel). Protein loading was controlled with anti-Cdc19 Western blots (bottom panels in A–C).

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