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. 2013 Mar 1;24(5):566–577. doi: 10.1091/mbc.E12-07-0552

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© 2013 Richmond et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 6: — Slk19 protein forms dimers. (A) Both full-length and cleaved fragment of Slk19 physically interact with themselves. Diploid yeast strains containing either SLK19/SLK19-13Myc or SLK19-6HA/SLK19-13Myc were grown to log phase. The cells were harvested to prepare the whole cell extracts with a bead beater. The extracts were first incubated with anti-HA antibody, and then protein A/G-coated agarose beads were added. Western blotting was performed to detect Slk19-6HA and Slk19-13Myc in the whole-cell extracts and the immunoprecipitates. Asterisk indicates a nonspecific band. (B) Slk19 interacts with itself in vitro. Purified Slk19-HA-His was mixed with Slk19-His or Slk19-Myc-His, and then the mixtures were pulled down with anti-Myc antibody. Slk19 proteins with different tags in the mixture and immunoprecipitates were detected using Western blotting with anti-Myc and anti-HA antibodies. A similar pull-down assay was performed after the mixture of Slk19-Myc-His with Slk19-His or Slk19-HA-His. (C) The working model for Slk19-dependent kinetochore clustering and sister kinetochore cohesion.