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. Author manuscript; available in PMC: 2017 Jan 9.
Published in final edited form as: Mol Cell. 2015 Oct 15;60(2):220–230. doi: 10.1016/j.molcel.2015.09.017

Figure 3. RNA binding through RRM domain is critical for Whi3 phase separation.

Figure 3

(A) Schematics of Whi3 protein with a polyQ domain and an RNA recognition motif. Whi3 is predicted to be disordered in the polyQ region. (B) Schematics illustrate the mutated Whi3 constructs. In the no-RNA experiment (left column of images), salt concentration was lowered from 150mM to 75mM by adding buffer with no salt, final protein concentration was 5μM of Whi3 and for fragments and mutants the highest protein concentration each construct could be purified was used: 25 μM of whi3ΔpolyQ, 39μM of whi3-Y610A, 23 μM of whi3-F653A, 11 μM whi3-Y610-F653A, 23 μM Whi3ΔRRM. In the CLN3 mRNA experiment (right column of images), 7.4 μM protein at 150mM salt was used for all mutants and 53 nM CLN3 mRNA was added. All images were taken after 4 hours of either adding no-salt buffer or mRNA.