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. 2010 Dec 2;286(6):4620–4631. doi: 10.1074/jbc.M110.195313

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — Grr1 mediates degradation of processed Stp1 but not processed Stp2. A, expression of an N-terminal truncation mutant of Stp1 (Stp1ΔN-HA) or Stp2 (Stp2ΔN-HA) leads to constitutive activation of AGP1-lacZ expression. Wild-type (ZLY044) and stp1Δ stp2Δ double deletion mutant cells (ZLY827) carrying control vector pRS416, STP1ΔN-HA (pZL1635), or STP2ΔN-HA (pZL1637) plasmid were grown in SD medium ± leucine and β-galactosidase activities were assayed. B, a grr1Δ mutation partially stabilizes Stp1ΔN-HA, but not Stp2ΔN-HA. Wild-type (ZLY044) and grr1Δ mutant (ZLY1942) cells expressing STP1ΔN-HA or STP2ΔN-HA were grown in SD medium, and cycloheximide chase was carried out as described. C, quantitative analysis of Stp1ΔN-HA and Stp2ΔN-HA levels in panel B. D, stabilization of Stp1ΔN-HA in grr1Δ cells leads to increased AGP1-lacZ expression. Cells described for panel B were grown in SD medium, and β-galactosidase activities were determined. Error bars represent S.E. of results from four independent experiments. The difference in AGP1-lacZ activity due to Stp1ΔN-HA expression in wild-type versus grr1Δ mutant cells is significant (p < 0.0001).