Identification of a DAF-16 Transcriptional Target Gene, scl-1, that Regulates Longevity and Stress Resistance in Caenorhabditis elegans Sadatsugu Ookuma,

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Identification of a DAF-16 Transcriptional Target Gene, scl-1, that Regulates Longevity and Stress Resistance in Caenorhabditis elegans Sadatsugu Ookuma, Makoto Fukuda, Eisuke Nishida Current Biology Volume 13, Issue 5, Pages 427-431 (March 2003) DOI: 10.1016/S0960-9822(03)00108-8

Figure 1 Identification of SCL-1 (A) One copy of DBE (DAF-16 consensus binding element) is located upstream of the first exon of scl-1. The DBE is boxed, and capital letters indicate the first methionine. (B) The predicted amino acid sequence of SCL-1. SCL-1 has a signal sequence and an SCP domain. The signal sequence is boxed, and the SCP domain is underlined. (C) A schematic of the domain structure of SCL-1 and its alignment with human CRISP family proteins. The percentages of the identity or the similarity to the SCP domain of SCL-1 are shown. Current Biology 2003 13, 427-431DOI: (10.1016/S0960-9822(03)00108-8)

Figure 2 Expression of scl-1 in Various Mutants and at Various Stages (A) Total RNA was isolated from stage-mixed cultures of each strain. scl-1 mRNA was upregulated in long-lived mutants (daf-2(e1370), age-1(hx546)) and was undetectable in daf-16(mgDf50) at 20°C. At 27°C, the level of scl-1 mRNA was increased in N2, daf-2(e1370), and age-1(hx546), but it was undetectable in daf-16(mgDf50). (B) Total RNA was isolated from synchronized cultures of each strain. Current Biology 2003 13, 427-431DOI: (10.1016/S0960-9822(03)00108-8)

Figure 3 Suppression of the Life Extension Phenotype of daf-2 by scl-1 RNAi Treatment (A) The level of scl-1 mRNA was greatly reduced in a scl-1 RNAi-treated daf-2(e1370) mutant at 20°C. (B) The life extension of daf-2(e1370) was significantly suppressed by scl-1 RNAi treatment, and the life span of N2 was slightly but reproducibly reduced by the treatment. The sums of two or more replications are shown. Mean life span ± SEM: daf-2(e1370), 26.5 ± 0.7 days (n = 128); daf-2; scl-1(RNAi), 19.0 ± 0.8 days (n = 137, p < 0.001); N2, 14.1 ± 0.6 days (n = 144); N2; scl-1(RNAi), 12.9 ± 0.4 days (n = 130, p = 0.13). Mean life spans of non-RNAi-treated animals, daf-2(e1370) and N2, on HT115 bacteria were not statistically different from those on OP50. (C) Dark intestinal pigmentations were reduced in the scl-1 RNAi daf-2(e1370). daf-2(e1370) (12 days after L4); daf-2; scl-1(RNAi) (12 days after L4); N2 (12 days after L4). (D) Autofluorescence in 5-day- and 10-day-old adult daf-2(e1370), daf-2; scl-1(RNAi), and N2 animals. (E) Intestine of adult daf-2(e1370), daf-2; scl-1(RNAi), and N2 animals. Current Biology 2003 13, 427-431DOI: (10.1016/S0960-9822(03)00108-8)

Figure 4 scl-1 Regulates Stress Resistance (A) scl-1 mRNA was upregulated after exposure to three stressers, heat (27°C or 35°C, 2 hr), UV light (0.2 J), and starvation (12 hr) in N2. (B) The RNAi treatment for scl-1 decreased resistance against heat shock (35°C) in N2. The sums of two or more replications are shown. Mean surviving life span ± SEM: N2, 14.6 ± 0.5 hr (n = 54); N2; scl-1(RNAi), 9.8 ± 0.1 hr (n = 54, p < 0.001). (C) The RNAi treatment for scl-1 decreased resistance against UV light (0.2 J) in N2. Mean surviving life span ± SEM: N2, 3.5 ± 0.1 days (n = 60); N2; scl-1(RNAi), 2.7 ± 0.1 days (n = 60, p = 0.007). Mean survival spans of N2 on HT115 were not statistically different from those on OP50 in (B) and (C). Current Biology 2003 13, 427-431DOI: (10.1016/S0960-9822(03)00108-8)