Light-sensitive phosphorylation regulates retinal IMPDH1 activity and filament assembly.

Inosine monophosphate dehydrogenase (IMPDH) is the rate-limiting enzyme in guanosine triphosphate (GTP) synthesis and assembles into filaments in cells, which desensitizes the enzyme to feedback inhibition and boosts nucleotide production. The vertebrate retina expresses two splice variants IMPDH1(546) and IMPDH1(595). In bovine retinas, residue S477 is preferentially phosphorylated in the dark, but the effects on IMPDH1 activity and regulation are unclear. Here, we generated phosphomimetic mutants to investigate structural and functional consequences of S477 phosphorylation. The S477D mutation resensitized both variants to GTP inhibition but only blocked assembly of IMPDH1(595) filaments. Cryo-EM structures of both variants showed that S477D specifically blocks assembly of a high-activity assembly interface, still allowing assembly of low-activity IMPDH1(546) filaments. Finally, we discovered that S477D exerts a dominant-negative effect in cells, preventing endogenous IMPDH filament assembly. By modulating the structure and higher-order assembly of IMPDH, S477 phosphorylation acts as a mechanism for downregulating retinal GTP synthesis in the dark when nucleotide turnover is decreased.

Light-sensitive phosphorylation regulates enzyme activity and filament assembly of human IMPDH1 retinal splice variants.

Inosine monophosphate dehydrogenase (IMPDH) is the rate-limiting enzyme in de novo guanosine triphosphate (GTP) synthesis and is controlled by feedback inhibition and allosteric regulation. IMPDH assembles into micron-scale filaments in cells, which desensitizes the enzyme to feedback inhibition by GTP and boosts nucleotide production. The vertebrate retina expresses two tissue-specific splice variants IMPDH1(546) and IMPDH1(595). IMPDH1(546) filaments adopt high and low activity conformations, while IMPDH1(595) filaments maintain high activity. In bovine retinas, residue S477 is preferentially phosphorylated in the dark, but the effects on IMPDH1 activity and regulation are unclear. Here, we generated phosphomimetic mutants to investigate structural and functional consequences of phosphorylation in IMPDH1 variants. The S477D mutation re-sensitized both variants to GTP inhibition, but only blocked assembly of IMPDH1(595) filaments and not IMPDH1(546) filaments. Cryo-EM structures of both variants showed that S477D specifically blocks assembly of the high activity assembly interface, still allowing assembly of low activity IMPDH1(546) filaments. Finally, we discovered that S477D exerts a dominant-negative effect in cells, preventing endogenous IMPDH filament assembly. By modulating the structure and higher-order assembly of IMPDH, phosphorylation at S477 acts as a mechanism for downregulating retinal GTP synthesis in the dark, when nucleotide turnover is decreased. Like IMPDH1, many other metabolic enzymes dynamically assemble filamentous polymers that allosterically regulate activity. Our work suggests that posttranslational modifications may be yet another layer of regulatory control to finely tune activity by modulating filament assembly in response to changing metabolic demands.

How Inhibitory Control Relates to Positive and Negative Affective States in Red Junglefowl.

Individual differences in inhibitory control, an aspect of cognition, are found in many species. How this variation links to affective states is not much explored, and could be relevant for welfare. As less fearful, more optimistic, individuals may act more impulsively, inhibitory control could link to less negative, more positive, affective states. Alternatively, poorer inhibitory control could associate with more negative, less positive, affective states, as poorer inhibitory control can result in individuals being less able to adapt to changing environments and more likely to show stereotypies. We here explored in three cohorts (N = 209) of captive red junglefowl, the ancestor of domestic chickens, how inhibitory control associated with affective states. Specifically, we measured inhibitory control with a detour task, and negative and positive affective states with a tonic immobility test and a cognitive judgement bias test, respectively. Cognition and behaviour can differ between ages and sexes. Therefore, we investigated how inhibitory control related to affective states in younger chicks (≈2.5 weeks old), older chicks (≈5 weeks old) and sexually mature adults (≈28 weeks old) of both sexes. In younger chicks, poorer inhibitory control associated with less negative, more positive, affective states. We found no relationship between inhibitory control and affective states in older chicks or adults, nor sex differences regarding how inhibitory control related to affective states. Overall, our results suggest that inhibitory control can link to affective states and that the nature of these links can change over ontogeny.