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. Author manuscript; available in PMC: 2017 Dec 5.
Published in final edited form as: Dev Cell. 2016 Nov 23;39(5):544–559. doi: 10.1016/j.devcel.2016.10.021

Figure 2. Development of conditional allele, sad1.2.

Figure 2

(A) Schematic of sad1 alleles with deletions in the nucleoplasmic domain. Transmembrane domain (TM) and SUN-domain are indicated. Each deletion allele replaced sad1+ and was GFP tagged. (B) The region subjected to PCR mutagenesis (2–60 aa) is indicated; see also Figure S3. (C) sad1.2 confers temperature sensitivity. Serial dilution (fivefold) of log-phase cultures (0.8 × 107 cells/mL) were spotted and grown on rich media. (D) Targeted mutations abolishing or mimicking phosphorylation at the two sites mutated in sad1.2 were used to replace wt sad1+ as in (A). Phosphorylation alone cannot account for sad1.2 temperature sensitivity.