Bi-directional ribosome scanning controls the stringency of start codon selection

Yifei Gu, Yuanhui Mao, Longfei Jia, Leiming Dong and Shu-Bing Qian ()
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Yifei Gu: Cornell University
Yuanhui Mao: Cornell University
Longfei Jia: Cornell University
Leiming Dong: Cornell University
Shu-Bing Qian: Cornell University

Nature Communications, 2021, vol. 12, issue 1, 1-12

Abstract: Abstract The fidelity of start codon recognition by ribosomes is paramount during protein synthesis. The current knowledge of eukaryotic translation initiation implies unidirectional 5ʹ→3ʹ migration of the pre-initiation complex (PIC) along the 5ʹ UTR. In probing translation initiation from ultra-short 5ʹ UTR, we report that an AUG triplet near the 5ʹ end can be selected via PIC backsliding. Bi-directional ribosome scanning is supported by competitive selection of closely spaced AUG codons and recognition of two initiation sites flanking an internal ribosome entry site. Transcriptome-wide PIC profiling reveals footprints with an oscillation pattern near the 5ʹ end and start codons. Depleting the RNA helicase eIF4A leads to reduced PIC oscillations and impaired selection of 5ʹ end start codons. Enhancing the ATPase activity of eIF4A promotes nonlinear PIC scanning and stimulates upstream translation initiation. The helicase-mediated PIC conformational switch may provide an operational mechanism that unifies ribosome recruitment, scanning, and start codon selection.

Date: 2021
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DOI: 10.1038/s41467-021-26923-3

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