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. 2007 Oct 8;27(24):8848–8858. doi: 10.1128/MCB.01186-07

FIG. 3.

FIG. 3.

The major sites of sumoylation in symplekin and CPSF-73 are conserved among species. (A) A plasmid encoding HA-SUMO-3 was cotransfected with plasmids encoding Flag-tagged wt symplekin or the indicated mutants into 293T cells, and lysates subjected to immunoblotting with anti-Flag antibodies. The sumoylated forms of symplekin are indicated by arrows. The panel below is a CLUSTALW alignment of human, mouse, and Xenopus symplekin. The bottom panel shows an amino acid alignment of human symplekin and yeast Pta1. Conserved lysines are boxed. (B) 293T cells were cotransfected with plasmids encoding His-SUMO-3 and Flag-tagged wt CPSF-73 (lanes 2 and 12) or the indicated mutants. Lysates were prepared and subjected to immunoblotting with anti-Flag antibodies. Sumoylated forms of CPSF-73 are indicated by arrows. The bottom panel shows a CLUSTALW amino acid alignment of human, zebrafish, D. discoideum and S. cerevisiae homologs of CPSF-73. Conserved SUMO acceptor lysines are boxed. WB, Western blot; +, present; −, absent.