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. 2018 May;24(5):721–738. doi: 10.1261/rna.065037.117

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Published by Cold Spring Harbor Laboratory Press for the RNA Society

This is a work of the US Government.

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FIGURE 9. — Grc3 PNK active site motifs are critical for ribosome production in S. cerevisiae. Polysome profile analysis of transformed tetO₇-GRC3–Las1 5X Flag expressing WT and PNK variants (P-loop^K252A, P-loop^S253A, Walker B^D283A, Clasp^L361A/W364A, Lid^R433A/R440A) of Grc3. Black arrows mark ribosome halfmers and brackets highlight the polysomes. Sucrose gradient fractions were precipitated, run on SDS-PAGE gels, and analyzed by western blot with anti-Myc (Grc3), anti-Flag (Las1), and anti-RPL8 (60S ribosomal protein) antibodies.