A combinatorial cis-regulatory logic restricts color-sensing Rhodopsins to specific photoreceptor subsets in Drosophila

Color vision in Drosophila melanogaster is based on the expression of five different color-sensing Rhodopsin proteins in distinct subtypes of photoreceptor neurons. Promoter regions of less than 300 base pairs are sufficient to reproduce the unique, photoreceptor subtype-specific rhodopsin expression patterns. The underlying cis-regulatory logic remains poorly understood, but it has been proposed that the rhodopsin promoters have a bipartite structure: the distal promoter region directs the highly restricted expression in a specific photoreceptor subtype, while the proximal core promoter region provides general activation in all photoreceptors. Here, we investigate whether the rhodopsin promoters exhibit a strict specialization of their distal (subtype specificity) and proximal (general activation) promoter regions, or if both promoter regions contribute to generating the photoreceptor subtype-specific expression pattern. To distinguish between these two models, we analyze the expression patterns of a set of hybrid promoters that combine the distal promoter region of one rhodopsin with the proximal core promoter region of another rhodopsin. We find that the function of the proximal core promoter regions extends beyond providing general activation: these regions play a previously underappreciated role in generating the non-overlapping expression patterns of the different rhodopsins. Therefore, cis-regulatory motifs in both the distal and the proximal core promoter regions recruit transcription factors that generate the unique rhodopsin patterns in a combinatorial manner. We compare this combinatorial regulatory logic to the regulatory logic of olfactory receptor genes and discuss potential implications for the evolution of rhodopsins. Author summary Each type of sensory receptor neuron in our body expresses a specific sensory receptor protein, which allows us to detect and discriminate a variety of environmental stimuli. The regulatory logic that controls this spatially precise and highly restricted expression of sensory receptor proteins remains poorly understood. As a model system, we study the mechanisms that control the expression of different color-sensing Rhodopsin proteins in distinct subtypes of Drosophila photoreceptors, which is the basis for color vision. Compact promoter regions of less than 300 base pairs are sufficient to reproduce the non-overlapping rhodopsin patterns. However, the regulatory logic that underlies the combination (sometimes called 'grammar') of the cis-regulatory motifs (sometimes called 'vocabulary') within the rhodopsin promoters remains poorly understood. Here, we find that specific combinations of cis-regulatory motifs in the distal and the proximal core promoter regions of each rhodopsin direct its unique expression pattern.

Automated summary

This study investigates the regulation of color-sensing Rhodopsin proteins in Drosophila photoreceptors, suggesting that both distal and proximal core promoter regions play a role in generating unique expression patterns. The proximal core promoter regions are not only involved in general activation but also contribute to generating non-overlapping expression patterns. Combinatorial cis-regulatory motifs in both promoter regions recruit transcription factors to generate the distinct rhodopsin patterns.