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Figure 5


Fig. 5. Lsm8p nuclear localization is affected by C-terminal, N-terminal and Sm-site deletions. (A) Cartoon showing the structures of the Lsm8 deletion constructs that were tested as GFP-fusion proteins. (B) Localization of GFP-tagged Lsm8p constructs in yeast strain MPS26 plus (i) pMPS8 (GFP-Lsm8), (ii) pMPS8-82 (GFP–Lsm8-2), (iii) pMPS8-313 (GFP–Lsm8-313), (iv) pMPS8C (GFP-Lsm8Cterm), (v) pMPS8-6A3 (GFP–Lsm8-6A3), (vi) pMPS8-{Delta}N (GFP-Lsm8{Delta}N), (vii) pMPS8-{Delta}SM (GFP-Lsm8{Delta}Sm); or in (vii) BMA38a plus pMPS8 (i.e. GFP-Lsm8 produced in the presence of endogenous Lsm8p) or (viii) pGFP-N-FUS (GFP). In all cases, GFP localization was examined in live cells after growth in SD-Ura-Met (to induce expression of the GFP construct). Bar, 10 µm. (C) Co-immunoprecipitation of myc-tagged Lsm7p with GFP-Lsm8p. Lsm8p-containing complexes were precipitated with anti-GFP antibodies, and Lsm7p was detected with anti-myc antibodies. (D) All the GFP-Lsm8 constructs were stably expressed, as shown by western blotting (and data not shown) using anti-GFP antibodies.





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