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. 2012 Oct 18;41(1):264–276. doi: 10.1093/nar/gks958

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© The Author(s) 2012. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial reuse, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com.

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Figure 8. — eEF2/GTP/SOR or eEF2/GTP/FA inhibits ribosome splitting but not release of mRNA. (A–E) eEF3/ATP releases 80S ribosomes from the PoTC in the presence of eEF2/GTP/SOR or FA. (A) No additions. (B) ATP/PUR was added. (C) Complete reaction mix for disassembly. (D) Inhibitor mix (eEF2/GTP/SOR) was added to C. (E) Inhibitor mix (eEF2/GTP/FA) was added to C. Note the large peak of 80S in D and E (arrows), but the amount of polysomes are about the same for these three conditions. (F and G) Lack of incorporation of externally added labelled 60S subunits into 80S ribosomes formed from PoTC by eEF3/ATP in the presence of eEF2/GTP/SOR or FA. The reaction shown in Table 1 with 15 pmol of PoTC and other components was performed in the presence of ³²P-labelled 60S subunits (0.25 pmol) with eEF2/GTP/SOR (F) or FA (G). The mixture was subjected to 5–30% SDGC, and both UV absorbance and radioactivity were measured for each fraction.