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. 2016 Feb 26;7(2):93–106. doi: 10.1080/21541248.2016.1156803

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© 2016 The Author(s). Published with license by Taylor & Francis Group, LLC

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

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Figure 4. — Results of the yeast two-hybrid screens with Sec4. (A) All 10 putative targets from the pull-down experiments were cloned into bait-vectors and tested for auto-activation against empty prey-vectors. In these experiments, only Mrs6 displayed auto-activation. (B) The Y2H experiments were repeated using the prey-vectors containing Sec4_WTΔC, Sec4_S34NΔC, Sec4_Q79LΔC or Sec4_D136NΔC to test for interaction with putative binding partners. Growth on the selective medium lacking histidine (-His) indicates an interaction of both known GEFs (Sec2 and Dss4) with Sec4_S34NΔC and Sec4_D136NΔC, but not with Sec4_WTΔC and Sec4_Q79LΔC. However the interaction is probably weak (as expected for an enzymatic interaction) and turnover of the chromogenic substrate X-α-Gal could only be observed for Dss4, but not Sec2 ((+) and (−): positive (pGBKT7–53/pGADT7-T) and negative (pGBKT7-Lam/pGADT7-T) controls).

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