Figure 5
From: Spam1-associated transmission ratio distortion in mice: Elucidating the mechanism
Spam1 RNA contains four AREs in the 3' UTR and UV-cross-linking reveals that one or two specifically bind testicular proteins. A) Four AU-rich motifs, 5' AUUUG...AUUUUA...AUUUUUA...AUUUUUG...3', are found in a 77 nucleotide sequence (nt 1909–1985) in Spam1 3' UTR. B) In the upper panel the thin line represents the 77 nt riboprobe sequence containing the AU motifs while the thick lines show the antisense DNA oligomers used in competition assays. C1 contains antisense oligos for all four AREs, C2 for the two at the 5' end of the sequence, C3 for the two at the 3' end, C5for the third from the 5' end, and C6 for the 3' end motif. The lower panel shows the results of in vitro label transfer by UV cross-linking and the RNA-protein complexes formed with the riboprobe and testicular protein extract, without competition, in Lanes T1 and T3. Lanes C1 and C2 show the disappearance of the complexes after competition with antisense oligos for all four motifs and the two at the 5' end, respectively. Lane C3 shows that the complexes are not diminished with competition with antisense oligos for the two 3' end AREs.