Abstract
Juvenile CLN3 disease is a recessively inherited paediatric neurodegenerative disorder, with most patients homozygous for a 1-kb intragenic deletion in CLN3. The btn1 gene is the Schizosaccharomyces pombe orthologue of CLN3. Here, we have extended the use of synthetic genetic array (SGA) analyses to delineate functional signatures for two different disease-causing mutations in addition to complete deletion of btn1. We show that genetic-interaction signatures can differ for mutations in the same gene, which helps to dissect their distinct functional effects. The mutation equivalent to the minor transcript arising from the 1-kb deletion (btn1102–208del) shows a distinct interaction pattern. Taken together, our results imply that the minor 1-kb deletion transcript has three consequences for CLN3: to both lose and retain some inherent functions and to acquire abnormal characteristics. This has particular implications for the therapeutic development of juvenile CLN3 disease. In addition, this proof of concept could be applied to conserved genes for other mendelian disorders or any gene of interest, aiding in the dissection of their functional domains, unpacking the global consequences of disease pathogenesis, and clarifying genotype–phenotype correlations. In doing so, this detail will enhance the goals of personalised medicine to improve treatment outcomes and reduce adverse events.
Highlights
Juvenile CLN3 disease is a recessively inherited paediatric neurodegenerative disorder, with most patients homozygous for a 1-kb intragenic deletion in CLN3
We have developed yeast models to understand the neuronal ceroid lipofuscinoses (NCL), known as Batten disease, a group of rare childhood inherited neurodegenerative disorders[1]
Each Bioneer library mutant was crossed with each query strain independently 3 times and pinned in quadruplicate, providing a total of 12 replicate colonies for each double mutant (Fig. 1A)
Summary
Juvenile CLN3 disease is a recessively inherited paediatric neurodegenerative disorder, with most patients homozygous for a 1-kb intragenic deletion in CLN3. We extended application of the powerful synthetic genetic array technique using Schizosaccha‐ romyces pombe to perform three independent systematic screens for three mutant strains of the orthologue of CLN3, btn[1] These strains were (1) btn[1] encoding a mutation mimicking one transcriptional effect of the 1-kb deletion (btn1102–208del)[6,7], (2) btn[1] encoding the equivalent of disease-associated missense mutation p.Asp416Gly (btn1D363G)[6], (3), and complete deletion of btn[1] (btn1Δ)[8]. Does not cause complete loss of function, but produces a complex partially functional Btn[1] protein with a unique gain of new characteristics This approach could be extended to understand functional consequences of diseaseassociated alleles in other genes with yeast orthologues, in addition to more generally dissecting the functional domains in any protein of interest
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