Abstract
The genetics underlying autism spectrum disorder (ASD) are complex. Approximately 3–5% of ASD cases arise from maternally inherited duplications of 15q11.2-q13.1, termed Duplication 15q syndrome (Dup15q). 15q11.2-q13.1 includes the gene UBE3A which is believed to underlie ASD observed in Dup15q syndrome. UBE3A is an E3 ubiquitin ligase that targets proteins for degradation and trafficking, so finding UBE3A substrates and interacting partners is critical to understanding Dup15q ASD. In this study, we take an unbiased genetics approach to identify genes that genetically interact with Dube3a, the Drosophila melanogaster homolog of UBE3A. We conducted an enhancer/suppressor screen using a rough eye phenotype produced by Dube3a overexpression with GMR-GAL4. Using the DrosDel deficiency kit, we identified 3 out of 346 deficiency lines that enhanced rough eyes when crossed to two separate Dube3a overexpression lines, and subsequently identified IA2, GABA-B-R3, and lola as single genes responsible for rough eye enhancement. Using the FlyLight GAL4 lines to express uas-Dube3a + uas-GFP in the endogenous lola pattern, we observed an increase in the GFP signal compared to uas-GFP alone, suggesting a transcriptional co-activation effect of Dube3a on the lola promoter region. These findings extend the role of Dube3a/UBE3A as a transcriptional co-activator, and reveal new Dube3a interacting genes.
Highlights
Autism spectrum disorder (ASD) has a prevalence of 1 in 681, and is characterized by social communication deficits and repetitive behaviors[2]
Out of the 346 DrosDel lines, we found that Df(2 L)ED62, Df(2 L)ED105, and Df(2 R) ED2076 enhanced rough eyes when crossed to both GMR > Dube3a45 and GMR > Dube3a27
We used a combination of smaller deficiency lines eliminating smaller segments of the genome within each DrosDel line and uas-RNAi lines to identify the specific genes within each region that, when knocked down, enhanced rough eyes produced by Dube3a overexpression (Table 1)
Summary
Autism spectrum disorder (ASD) has a prevalence of 1 in 681, and is characterized by social communication deficits and repetitive behaviors[2]. While the underlying genetic causes of autism are complex, recurrent copy number variants associated with ASD can provide insight into the genetic mechanism as they are consistently observed at high rates among ASD individuals[3,4,5,6]. One such disorder results from maternally derived duplications of chromosome 15q11.2-q13.1, termed Duplication 15q (Dup15q) syndrome and the majority of Dup15q individuals meet the criteria for ASD7. We performed an enhancer/ suppressor screen, using a rough eye phenotype produced by over-expression of Dube3a with GMR-GAL4, to identify new Dube3a interacting genes. Using the DrosDel deficiency kit that covers 65.2% of the fly genome[13], and subsequent uas-RNAi lines, we found three new Dube3a interacting genes: GABA-B-R3, IA2, and lola
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