Allelic strengths of encephalopathy-associated UBA5 variants correlate between in vivo and in vitro assays

• DOI: 10.7554/elife.89891.3
• Publication Date: 2023-12-11T16:26:33Z
• Authors: Xueyang Pan; Albert N Alvarez; Mengqi Ma; Shenzhao Lu; Michael W Crawford; Lauren C Briere; Oguz Kanca; Shinya Yamamoto; David A Sweetser; Jenny L Wilson; Ruth J Napier; Jonathan N Pruneda; Hugo J Bellen
• Keywords: 0301 basic medicine; Medical Sciences; QH301-705.5; Science; allelic series; Mutation, Missense; 610; rare disease; Ubiquitin-Activating Enzymes; Article; Biomedical Informatics; 03 medical and health sciences; UBA5; Intellectual Disability; UFMylation; Medical Specialties; Medicine and Health Sciences; Drosophila Proteins; and Immunity; Humans; developmental and epileptic encephalopathy; Biology (General); Biological Phenomena; Brain Diseases; Movement Disorders; D. melanogaster; Cell Phenomena; Q; R; Life Sciences; Proteins; Genetics and Genomics; 3. Good health; Medical Molecular Biology; Mutation; Medicine; Drosophila; Missense; Medical Genetics; Human

Protein UFMylation downstream of the E1 enzyme UBA5 plays essential roles in development and endoplasmic reticulum stress. Variants in the UBA5 gene are associated with developmental and epileptic encephalopathy 44 (DEE44), an autosomal recessive disorder characterized by early-onset encephalopathy, movement abnormalities, global developmental delay, intellectual disability, and seizures. DEE44 is caused by at least 12 different missense variants described as loss of function (LoF), but the relationships between genotypes and molecular or clinical phenotypes remain to be established. We developed a humanized UBA5 fly model and biochemical activity assays in order to describe in vivo and in vitro genotype–phenotype relationships across the UBA5 allelic series. In vivo, we observed a broad spectrum of phenotypes in viability, developmental timing, lifespan, locomotor activity, and bang sensitivity. A range of functional effects was also observed in vitro across comprehensive biochemical assays for protein stability, ATP binding, UFM1 activation, and UFM1 transthiolation. Importantly, there is a strong correlation between in vivo and in vitro phenotypes, establishing a classification of LoF variants into mild, intermediate, and severe allelic strengths. By systemically evaluating UBA5 variants across in vivo and in vitro platforms, this study provides a foundation for more basic and translational UBA5 research, as well as a basis for evaluating current and future individuals afflicted with this rare disease.