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. 2019 Nov 19;48(2):862–878. doi: 10.1093/nar/gkz1092

Figure 3.

Figure 3.

MOV10 unwinds rG4, enhances rG4 luciferase activity from a reporter and enhances FMRP-mediated translation of a subset of RNAs through its N-terminus. (A) Purified murine MOV10 unwinds a synthesized rG4 reporter (iSpinach). Red: iSpinach in the presence of MOV10, DHFBI, in the absence of ATP. Blue: iSpinach in the presence of MOV10, DHFBI, and ATP. Green: MOV10 and ATP, absence of DFHBI. (B) Luciferase activity of MAZ 3′ UTR reporter in WT N2a, Mov10 KO N2a and Mov10 KO N2a transfected with the N-terminal MOV10 construct, (n = 3), error bars represent SD, ** P < 0.001 Student t-test. (C) Quantification of neurite length (μm) in WT or Mov10 KO N2a cells post differentiation transfected with null or N-,C-terminal MOV10, or a translocation mutant (K531A), error bars represent SEM, one way ANOVA (F(4.94) = 3.964, P = 0.0052, **P< 0.05,***P< 0.01. (D) Immunoblot of endogenous proteins encoded by mRNAs co-bound by FMRP–MOV10 in HEK293 cells expressing WT FMRP or FMRP-ΔRGG transfected with N-terminal MOV10. (Bottom) Quantification of protein levels, (n = 3), error bars represent SD, **P < 0.001 Student t-test. (E) RT-qPCR of IP’ed AGO-associated mRNAs in HEK293 cells expressing endogenous levels of MOV10 or over-expressing N-terminal MOV10, (n = 3), error bars represent SD, *P < 0.05, ** P < 0.001 Student t-test.