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The V48D and L125Q mutants promote strong 60S biogenesis and subunit-joining defects. Cytoplasmic extracts from isogenic strains were loaded onto 7–47% sucrose gradients, centrifuged in an SW41 rotor at 40 000 r.p.m. for 180 min at 4°C, fractionated and analyzed by continuous monitoring of A254 (48). The locations of 40S, 60S, 80S, polysome fractions and half-mers are labeled. The presence of half-mers is indicative of subunit-joining defects.
