Björn Schumacher, Kay Hofmann, Simon Boulton, Anton Gartner

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The C. elegans homolog of the p53 tumor suppressor is required for DNA damage- induced apoptosis Björn Schumacher, Kay Hofmann, Simon Boulton, Anton Gartner Current Biology Volume 11, Issue 21, Pages 1722-1727 (October 2001) DOI: 10.1016/S0960-9822(01)00534-6

Figure 1 (a) Predicted amino acid sequence of cep-1. (b) A profile-guided alignment of CEP-1 with other established members of the p53 family. Residues that interact with DNA in human p53 are marked with “D,” those involved in Zn binding are marked with “Z,” and those most frequently mutated in cancers are marked with an asterisk. The evolutionarily conserved regions II–IV and the tetramerization domain (tet) are indicated [16]. Abbreviations used are as follows: lolfo, Loligo forbes (northern European squid); mesau, Mesocricetus auratus (golden hampster); and myaar, Mya arenaria (mussle). (c) Neighbor-joining dendrogram of the p53 family. Overall, the dendrogram reflects the evolutionary relationships of the organisms involved. Obvious exceptions are p63 and p73, which have arisen from gene duplication events. CEP-1 robustly clusters with the Drosophila sequence (1000 out of 1000 bootstrap trials) Current Biology 2001 11, 1722-1727DOI: (10.1016/S0960-9822(01)00534-6)

Figure 2 CEP-1 is a transcription factor. (a) CEP-1 acts as a transcriptional activator. We introduced various Gal4 DNA binding domain fusions into yeast and measured transcriptional activation by determining β-Gal activity. (b) pSS1 p53 reporter construct. (c) Human and C. elegans p53 recognize the same consensus p53 binding site. p415ADH, p415ADH-BAR-1, p415ADH-human p53, and p415ADH-CEP-1 were introduced into yeast strains containing the pSS1 reporter (human p53 binding sites). Transcriptional activation of the reporter was determined by the ability of yeast strains to grow in the absence of histidine. A derivative of the pSS1 reporter construct lacking the p53 binding site did not support growth in the absence of histidine (S.B., unpublished observation) Current Biology 2001 11, 1722-1727DOI: (10.1016/S0960-9822(01)00534-6)

Figure 3 cep-1 is specifically required for DNA damage-induced apoptosis. (a) A Nomarski picture showing the pachytene section of the germline was taken 36 hr after 60 Gray irradiation. Corpses are indicated by arrows. Unrelated gfp sequence was used as control RNAi. (b) Germcell corpses were counted at different time points after irradiation. (c)cep-1RNAi phenotype is mimicked by cosuppression. 50 ng/μl cep-1 was coinjected with 50 ng/μl pRF-4 rol-6 marker. Germ cell apoptosis was scored 36 hr post irradiation Current Biology 2001 11, 1722-1727DOI: (10.1016/S0960-9822(01)00534-6)