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. 2009 May 2;284(27):17956–17967. doi: 10.1074/jbc.M109.013722

FIGURE 7.

FIGURE 7.

Drs2p-Cdc50p interactions are tied to the ATPase reaction cycle. A, schematic representation of P2A and P4-ATPase reaction cycles. Residues important for E1 → E1P, E1P → E2P, and E2P → E2 transitions in the P2A ATPase SERCA1a and the corresponding residues in the P4-ATPase Drs2p are indicated (⊥). B, split ubiquitin growth assay reporting DRS2-Cub, DRS2D560N-Cub, DRS2E342Q-Cub, and DRS2G341L-Cub interactions with Nub-CDC50 subunits. C, modulation of Cub fusion expression by methionine was performed as in Fig. 6C. D, split ubiquitin β-galactosidase (β-gal.) assay, performed as in Fig. 6D. Data shown are representative of two independent experiments. E, mutation of Asp-560, Gly-341, or Glu-342 causes loss of Drs2p function in vivo. Serial dilutions of a Δdrs2 mutant strain transfected with pEV-Cub (empty vector (EV) control), pDRS2-Cub, pDRS2D560N–Cub, pDRS2G341L–Cub, or pDRS2E342Q were spotted onto SD plates and incubated at 30 or 18 °C for 4 days.