JCS002808 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 -

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• Supplemental Figure S1 -

  Fig. S1. The S103P mutation activates the Ire1 core mutant. Left, the luminal domain of the mutated Ire1 is illustrated as in Fig. 1A. The positions of five point mutations introduced according to Papa et al. (2003) are indicated. The 5m mutant carried all five substitutions (S103P, E167D, Q272R, F377S and H446N), whereas the other mutants had one or more of the five possible substitutions. Right, KMY1015 (ire1 null mutant) cells carrying both the UPRE-lacZ reporter pCZY1 and a mutant allele of pRS315-Ire1-HA were cultured without extrinsic stress, and subjected to β-galactosidase assay. Each value is the mean ± s.d. of three independent clones and was normalized to that of the Tun⁺ wild-type Ire1-HA control, which was set at 100.

• Supplemental Figure S2 -

  Fig. S2. Activity of the Ire1 mutants that carry no epitope tag. KMY1015 (ire1 null mutant) cells carrying both the UPRE-lacZ reporter pCZY1 and a wild-type or mutant allele of pRS313-Ire1 were cultured with (+) or without (−) 2 μg/ml tumicamycin (Tun) for 4 hours, and assayed for β-galactosidase. Each value is the mean ± s.d. for three independent clones and was normalized to that of the Tun⁺ wild-type control (untagged), which was set at 100.

• Supplemental Figure S3 -

  Fig. S3. A control experiment for Fig. 5A. An experiment similar to that shown in Fig. 5A was performed using empty vector pRS315 instead of derivatives of the Ire1-HA plasmid pRS315-Ire1-HA.