Ribosomal mutation in helix 32 of 18S rRNA alters fidelity of eukaryotic translation start site selection

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 eIF5^G31R 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 eIF5^G31R mutant. The C1209U mutation suppressed Sui^- and Gcd^- (constitutive derepression of GCN4 expression) phenotype of eIF2β^S264Y , eIF1^K60E , and eIF1A-ΔC mutation. We propose that the C1209U mutation in 40S ribosomal may perturb the premature head rotation in 'Closed/P_IN ' state and enhance the stringency of translation start site selection.