The 40S ribosome plays a critical role in start codon selection. To gain insights into the role of its 18S rRNA in start codon selection, a suppressor screen was performed that suppressed the preferential UUG start codon recognition (Suppressor of initiation codon: Sui− phenotype) associated with the eIF5G31R mutant. The C1209U mutation in helix h32 of 18S rRNA was found to suppress the Sui− and Gcn− (failure to derepress GCN4 expression) phenotype of the eIF5G31R mutant. The C1209U mutation suppressed Sui− and Gcd− (constitutive derepression of GCN4 expression) phenotype of eIF2βS264Y, eIF1K60E, and eIF1A‐ΔC mutation. We propose that the C1209U mutation in 40S ribosomal may perturb the premature head rotation in ‘Closed/PIN’ state and enhance the stringency of translation start site selection.
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