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. 1999 Nov 9;96(23):13208–13213. doi: 10.1073/pnas.96.23.13208

Figure 1.

Figure 1

In vivo binding of Bir1p to Ndc10p. (A) Two-hybrid interaction between a Bir1p fragment and known kinetochore proteins. A tester strain PJ69–4A (17) carrying the plasmid pACD-Bir1 (C332) was transformed with various plasmids that express DBD alone or DBD-fused Ndc10p, Cep3p, Ctf13p, Skp1p, or Cbf1p. The transformants were streaked on SD/His− medium containing 2 mM 3-AT (3-amino-1 2,4-triazole) to test expression of the HIS3 reporter gene. (B) Two-hybrid assay with strain PJ69-4A carrying pDBD-Bir1 (full). Strain PJ69-4A/pDBD-Bir1 (full) bearing plasmids expressing ACD-fused Mif2p, Cse4p, Ctf19p, Mcm21p, Okp1p, or Slk19p did not grow on minimal salt-dextrose/His− medium containing 2 mM 3-AT (not shown). (C) Identification of Bir1p regions interacting with Ndc10p. (Top) Schematic view of Bir1p showing the relative locations of BIR domains 1 (R1), 2 (R2), and of nuclear localization signal (NLS), a bipartite nuclear localization signal. Various truncated Bir1p fragments as shown here (See Materials and Methods) were tested for two-hybrid interactions with Ndc10p on SD/His− medium containing 2 mM 3-AT.