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. 2013 Jun 22;10:205. doi: 10.1186/1743-422X-10-205

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Copyright © 2013 Morais et al.; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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The mapping of the terminal region residues of the interaction domain using the yeast two-hybrid system indicates a critical interaction with eIF3L. The yeast was co-transformed with the mutant pGBKT7 constructs and pACT2-eIF3L. A positive interaction is shown by the activation of the reporter gene HIS3 in the interaction of eIF3L with the mutants ID (D436N), ID (D436S), and ID (R439A/H442A), indicating that the mutations are not critical for the interaction. A negative interaction between mutant ID (F431A/W432A/V435A) and eIF3L is indicated by the absence of growth on plates lacking histidine (−His). C-, negative control; C+, positive control; N, normalized original culture sample; 1–4, 10-fold serial dilutions of sample.