FIGURE 1: Identification of human TC-1 as a Bax suppressor that protects against programmed cell death-inducing levels of copper.

(A) Plasmid DNA with the Bh1 sequence was obtained from the clone identified from the original cDNA library screen for Bax suppressors [52]. It was then reintroduced into naïve BY4742 yeast cells with and without Bax. A human 14-3-3β/α cDNA was used as a positive control [53]. Cultures of the transformants were grown, serially diluted, and spotted onto YNB media with glucose or with galactose. The plates were allowed to grow at 30°C and photographs of the plates are shown.

(B) The amino acid sequence of human TC-1 was used to search the available protein databases at NCBI. Boxshade of the TCoffee alignment of representative sequences from different species showing strong similarity with hTC-1 are shown. The black shading indicates identical residues while grey indicates conserved residues. The scientific names and database accession numbers of the species used are as follows: human: Homo sapiens (Accession Number NM_020130); cow: Bos taurus (Accession Number NP_001030567); mouse: Mus musculus (Accession Number NP_081207); python: Python bivittatus (Accession Number XP_007430414.1); pigeon: Columba livia (Accession Number XP_005510897.1); frog: Xenopus tropicalis (Accession Number XP_002936483.2); fish: Poecilia formosa (Accession Number XP_007573497.1); and shark: Callorhinchus milii (Accession Number XP_007883063.1).

52. Yang Z, Khoury C, Jean-Baptiste G, Greenwood MT (2006). Identification of mouse sphingomyelin synthase 1 as a suppressor of Bax-mediated cell death in yeast. FEMS Yeast Res 6(5): 751-762.

53. Clapp C, Portt L, Khoury C, Sheibani S, Norman G, Ebner P, Eid R, Vali H, Mandato CA, Madeo F, Greenwood MT (2012). 14-3-3 Protects against stress-induced apoptosis. Cell Death Dis 3:e348.

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