Magenta arrowheads = additional serines mutated in 4 serine and six serine mutations (data not shown).
We sequenced all the coding exons and flanking regions of NIN gene but did not reveal any other pathogenic mutations (data not shown).
Meanwhile, similar to the LSE1 mutant line, Southern blot analysis revealed that neither the LSE2 nor the LSE3 phenotype was tagged by the Tos17 retrotransposon, indicating that these phenotypes were caused by some other mutations (data not shown).
Tagged SUMO rescues the lethality resulting from sumo mutations (data not shown) demonstrating the functionality of the tagged protein.
PTEN exon 8 was further evaluated by reverse direction Sanger sequencing which also revealed no mutations (data not shown).
The mre11 gene duplication is estimated to be about 15 million years old, under the assumption of a molecular clock for synonymous mutations (data not shown).
This cav gene duplication is estimated to be 10 million years old, under the assumption of a molecular clock for synonymous mutations (data not shown).
An independent hos2Δ/Δ start-to-stop deletion mutant also showed enhanced filamentation, corroborating the results of the insertional mutations (data not shown).
All have seemingly been inactive for a considerable period as suggested by their long terminal lineages plus numerous indels and non-sense mutations (data not shown).
We found that pmk-3 mutations did not enhance the phenotype of unc-11 mutations (data not shown), suggesting that mutations in unc-11 can occlude the effect of mutations in pmk-3.
Indeed, this cav gene duplication is estimated to be as old as the split between the Sophophora and Drosophila subgenera, and thus about 40 million years old, under the assumption of a molecular clock for synonymous mutations (data not shown).
