Non-optimal codon usage affects expression, structure and function of clock protein FRQ. Nature 2013

Codon usage bias has been observed in almost all genomes and is thought to result from selection for efficient and accurate translation of highly expressed genes1–3. Codon usage is also implicated in the control of transcription, splicing and RNA structure4–6. Many genes exhibit little codon usage bias, which is thought to reflect a lack of selection for mRNA translation. Alternatively, however, non-optimal codon usage may have biological significance. The rhythmic expression and the proper function of the Neurospora FREQUENCY (FRQ) protein are essential for circadian clock function. Here we show that, unlike most genes in Neurospora, frq exhibits non-optimal codon usage across its entire open reading frame. Optimization of frq codon usage abolishes both overt and molecular circadian rhythms. Codon optimization not only increases FRQ level but surprisingly, also results in conformational changes in FRQ protein, altered FRQ phosphorylation profile and stability, and impaired functions in the circadian feedback loops. These results indicate that non-optimal codon usage of frq is essential for its circadian clock function. Our study provides an example of how non-optimal codon usage functions to regulate protein expression and to achieve optimal protein structure and function