Figure 5.

Tdrd6a and Buc Interact via sDMAs in the C Terminus of Buc

(A) Volcano plot of Tdrd6a IP compared to IgG IP on embryo extracts, followed by MS.

(B) Confirmation of Tdrd6a co-IP with Buc using the Buc-eGFP transgenic line.

(C) C terminus of Buc with the identified dimethylated peptide underlined. Asterisk indicates residues that were found to be dimethylated by MS. Three RG sites together form a tri-RG motif, indicated in gray.

(D) Volcano plot of peptide pull-down on embryo extracts followed by MS. On the “Methylated” peptide, all 3 RG motifs were symmetrically dimethylated.

(E) Peptide pull-down followed by western blot for multiple methylated (sDMA) and non-methylated peptides derived from proteins known to contain sDMA modifications and the Buc homolog XVelo on ovary extracts. Listed are all peptides used.

(F) Peptide pull-down of Buc C terminus peptides with different methylation states on embryo extracts.

(G) MS of pull-downs of double and triple sDMA modified peptides.

(H) MS of Tdrd6a IP in the buc^+/− compared to buc^−/− background. Tdrd6c and Ziwi are specifically enriched in the buc^+/− background, indicating that they require the presence of Buc to associate with Tdrd6a.