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. 2012 May 4;31(11):2461–2472. doi: 10.1038/emboj.2012.102

Figure 2.

Figure 2

Sumoylation of Kap114 is required for its proper cellular localization. (A) Kap114 K909R accumulates in the nucleus. Δkap114 cells containing pRS316–KAP114–GFP or pRS316–KAP114 K909R–GFP were cultured in liquid medium at 30°C. (B) The mutagenesis of the ΨKxD/E sumoylation consensus motif between residues 908 and 911 affects the cellular distribution of Kap114. Cells deleted for KAP114, transformed with pRS316–KAP114–GFP plasmids, and carrying the unmutated allele or the indicated KAP114 mutations were grown in liquid medium. (C) WT cells, Δulp2, ulp1–333, or uba2–ts9 mutants containing the plasmids pRS316–KAP114–GFP, pRS316–GFP–KAP95, or pRS316–GFP–CSE1 were cultured in liquid media at 30°C (WT and Δulp2) or 25°C (ulp1–333 and uba2–ts9). Logarithmically growing cells were analysed by direct fluorescence microscopy using a Zeiss Axioscope microscope at 1000-fold magnification to determine the localization of the GFP fusion proteins. Scale bar, 5 μm.