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. 2010 Jul 15;466(7304):383-7.
doi: 10.1038/nature09195. Epub 2010 Jun 16.

Members of the H3K4 Trimethylation Complex Regulate Lifespan in a Germline-Dependent Manner in C. Elegans

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Free PMC article

Members of the H3K4 Trimethylation Complex Regulate Lifespan in a Germline-Dependent Manner in C. Elegans

Eric L Greer et al. Nature. .
Free PMC article

Abstract

The plasticity of ageing suggests that longevity may be controlled epigenetically by specific alterations in chromatin state. The link between chromatin and ageing has mostly focused on histone deacetylation by the Sir2 family, but less is known about the role of other histone modifications in longevity. Histone methylation has a crucial role in development and in maintaining stem cell pluripotency in mammals. Regulators of histone methylation have been associated with ageing in worms and flies, but characterization of their role and mechanism of action has been limited. Here we identify the ASH-2 trithorax complex, which trimethylates histone H3 at lysine 4 (H3K4), as a regulator of lifespan in Caenorhabditis elegans in a directed RNA interference (RNAi) screen in fertile worms. Deficiencies in members of the ASH-2 complex-ASH-2 itself, WDR-5 and the H3K4 methyltransferase SET-2-extend worm lifespan. Conversely, the H3K4 demethylase RBR-2 is required for normal lifespan, consistent with the idea that an excess of H3K4 trimethylation-a mark associated with active chromatin-is detrimental for longevity. Lifespan extension induced by ASH-2 complex deficiency requires the presence of an intact adult germline and the continuous production of mature eggs. ASH-2 and RBR-2 act in the germline, at least in part, to regulate lifespan and to control a set of genes involved in lifespan determination. These results indicate that the longevity of the soma is regulated by an H3K4 methyltransferase/demethylase complex acting in the C. elegans germline.

Figures

Fig. 1
Fig. 1. The ASH-2, WDR-5, and SET-2 function together to regulate H3K4me3 and lifespan in C. elegans
a, Percent change in average lifespan in worms treated with RNAi to the indicated genes compared with empty vector. *: p<0.05. b, Western blots on L3 worm extracts (representative of 4 independent experiments). c, wdr-5/tag-125(ok1417) mutant worms have an extended lifespan. d, set-2 knock-down extends lifespan. e, Western blot of L3 worm extracts after SET-2 knock-down or deletion (representative of 3 independent experiments). f, The methyltransferase domain of worm SET-2 (SET-2SET) methylates histone H3 in vitro. e, ash-2 and wdr-5/tag-125 knock-down slightly extend the lifespan of set-2(ok952) mutant worms, but significantly less than in the WT (N2) background. ash-2 mRNA was efficiently knocked-down by RNAi in set-2(ok952) mutant worms (Supplementary Fig. 3a). E.V.: empty vector. Statistics are presented in Supplementary Tables 1 and 2.
Fig. 2
Fig. 2. RBR-2 is an H3K4me3 demethylase that counteracts the effect of the ASH-2 methyltransferase complex
a, Western blot of rbr-2(tm1231) mutant worms (representative of 3 independent experiments). b, rbr-2(tm1231) mutant worms have a decreased lifespan. c, Western blots of rbr-2(tm1231) mutant worms treated with ash-2 RNAi (representative of 2 independent experiments). d, ash-2 knock-down does not extend the lifespan of rbr-2(tm1231) mutant worms. ash-2 mRNA was efficiently knocked-down by RNAi in rbr- 2(tm1231) mutant worms (Supplementary Fig. 3a). E.V.: empty vector. Statistics are presented in Supplementary Table 3.
Fig. 3
Fig. 3. An intact germline is necessary for longevity and gene expression control by the H3K4me3 regulatory complex
a, b, Whole-mount immunofluorescence of young adult worms stained with an ASH-2 antibody (a) or an H3K4me3 antibody (b). Dashed lines marks the germline. Scale bars: 50 μm. c, ash-2 knock-down does not further extend the long lifespan of glp-1(e2141ts) mutant worms that were shifted to the restrictive temperature at the L1 stage. d, set-2 knock-down does not further extend the long lifespan of germline-deficient glp-1(e2141ts) mutant worms that were shifted to the restrictive temperature at the L1 stage. Statistics are presented in Supplementary Table 4. e, Microarray clusters of genes that change significantly upon ash-2 knock-down in wildtype worms, but not in glp-1(e2141ts) mutants. Experimental values are presented in Supplementary Tables 8 and 9. f, Percentage of genes regulated by ASH-2 only in WT worms but not in glp-1(e2141ts) mutants. g, Top gene ontology (GO) terms for genes regulated by ASH-2 in wildtype worms, but not in glp-1(e2141ts) mutants. E.V.: empty vector.
Fig. 4
Fig. 4. ASH-2/RBR-2 function primarily in the germline to regulate lifespan and require the continuous production of mature eggs for lifespan extension
a, ash-2 RNAi extends the lifespan of rrf-1(pk1417) mutant worms, which are deficient for RNAi in the soma, to a similar extent as in WT (N2) worms. b, Three independent lines of low-copy (LC) integrant Ppie-1::rbr-2::gfp transgenic worms have increased lifespan compared to two independent lines of Ppie-1::gfp transgenic worms. c, ash-2 knock-down does not further extend the long lifespan of glp-1(e2141ts) mutant worms that were shifted to the restrictive temperature at the young adult stage. d, ash-2 knockdown does not extend the lifespan of fem-3(e2006ts) mutant worms, which do not produce mature eggs. ash-2 mRNA was efficiently knocked-down by RNAi in fem-3(e2006ts) mutants (Supplementary Fig. 3b). E.V.: empty vector. Statistics are presented in Supplementary Tables 4 and 5.

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