4kb genomic fragment that contains the Sply gene, including 491bp of upstream regulatory sequences and the 3'UTR. The coding region has been mutated to contain the E119G amino acid replacement, which is equivalent to the disease-associated E132G mutation in the orthologous human SGPL1 gene. A Tag:HA tag has been inserted at the C-terminal end of the Sply open reading frame.
A17090174G
GAG>GGG
E119G | Sply-PA; E119G | Sply-PB
E119G
Analogous E132G mutation in human SGPL1 implicated in nephrotic syndrome, type 14; mutation carried on in vitro construct.
Either the SplyS335I.T:Ivir\HA1 or the SplyR210Q.T:Ivir\HA1 allele of Sply bearing mutations discovered to be disease-associated in its human ortholog SPGL1 fail to ameliorate the phenotype of Sply05091 mutants, the Drosophila model of nephrotic syndrome. The mutant phenotype can be rescued by combination with SplyE119G.T:Ivir\HA1.
In human, although this variant is predicted to result in a benign missense change, it appears to promote an abnormal upstream splice, which results in a frameshift.
The reduced density of foot process in garland cell nephrocytes observed in Sply05091/Df(2R)BSC433 third instar larvae is rescued by combination with SplyE119G.T:Ivir\HA1.
