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Figure 5


Fig. 5. Region III of Nvj1p is necessary and sufficient to bind Osh1p. (A) Deletion of the most conserved residues in region III abrogates the Nvj1p-dependent sequestration of GFP-Osh1p to the nuclear envelope. Full-length (FL) or N- and C-terminal truncations of Nvj1p were overexpressed in nvj1-{Delta} cells and tested for the ability to sequester GFP-Osh1p from soluble and Golgi-associated pools to the nuclear surface. Conserved residues in region III of Nvj1p (aa 130-176) proved necessary for GFP-Osh1p sequestration. Numbers correspond to amino acid positions. (B) Residues 130-177 of Nvj1p are sufficient to co-immunoprecipitate GFP-Osh1p in vivo. Lysates of cells expressing GFP-Osh1p and 3HA-tagged Nvj1p(130-177aa) or empty vector were prepared as described in Materials and Methods. Nvj1p(130-177aa)-3HA was immunoprecipitated (IP) with anti-HA conjugated agarose beads.