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Hypoxic response regulators RHY-1 and EGL-9/PHD promote longevity through a VHL-1-independent transcriptional response

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Abstract

HIF-1-mediated adaptation to changes in oxygen availability is a critical aspect of healthy physiology. HIF is regulated by a conserved mechanism whereby EGLN/PHD family members hydroxylate HIF in an oxygen-dependent manner, targeting it for ubiquitination by Von-Hippel-Lindau (VHL) family members, leading to its proteasomal degradation. The activity of the only C. elegans PHD family member, EGL-9, is also regulated by a hydrogen sulfide sensing cysteine-synthetase-like protein, CYSL-1, which is, in turn, regulated by RHY-1/acyltransferase. Over the last decade, multiple seminal studies have established a role for the hypoxic response in regulating longevity, with mutations in vhl-1 substantially extending C. elegans lifespan through a HIF-1-dependent mechanism. However, studies on other components of the hypoxic signaling pathway that similarly stabilize HIF-1 have shown more mixed results, suggesting that mutations in egl-9 and rhy-1 frequently fail to extend lifespan. Here, we show that egl-9 and rhy-1 mutants suppress the long-lived phenotype of vhl-1 mutants. We also show that RNAi of rhy-1 extends lifespan of wild-type worms while decreasing lifespan of vhl-1 mutant worms. We further identify VHL-1-independent gene expression changes mediated by EGL-9 and RHY-1 and find that a subset of these genes contributes to longevity regulation. The resulting data suggest that changes in HIF-1 activity derived by interactions with EGL-9 likely contribute greatly to its role in regulation of longevity.

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Acknowledgments

We thank David Angeles and the Sternberg lab for sharing their Kallisto analyses and expertise.

Funding

This work was supported by NIH R01AG058717 and the Glenn Foundation for Medical Research. HAM was supported by NIH F31AG060663. Additionally, the University of Washington Nathan Shock Center provided support of gene expression analysis (NIH P30AG013280). Strains were provided by the Caenorhabditis Genetics Center that is funded by the NIH ORIP (P40OD010440).

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Authors and Affiliations

  1. Molecular & Integrative Physiology Department, University of Michigan, Ann Arbor, MI, 48109, USA

    Joseph C.P. Kruempel, Megan L. Schaller, Abrielle Fretz, Marjana Sarker, Shijiao Huang & Scott F. Leiser

  2. Cellular and Molecular Biology Graduate Program, University of Michigan, Ann Arbor, MI, 48109, USA

    Hillary A. Miller & Marshall Howington

  3. Department of Internal Medicine, University of Michigan, Ann Arbor, MI, 48109, USA

    Scott F. Leiser

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  1. Joseph C.P. Kruempel
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Correspondence to Scott F. Leiser.

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Supplementary Figure 1.

rhy-1-mediated effects of lifespan. A-C) Individual lifespan experiments with N2 and rhy-1(ok1402) animals treated with empty vector or vhl-1(RNAi). D). Additional lifespan trial comparing N2 and rhy-1(ok1402) lifespans on empty vector RNAi. E. Aggregated data from A-C. (PNG 262 kb)

High resolution image (EPS 1119 kb)

Supplementary Figure 2.

Effects of additional VHL-1/EGL-9 antagonistic HIF-1 targets on lifespan. A, C) expression levels of selected transcripts from RNA-seq analyses. B-D) Treatment with ftn-1(RNAi), and ftn-2(RNAi) increases lifespan of egl-9(sa307);vhl-1(ok161) mutants (p < 0.05 by log-rank). Lifespan data are aggregated from at least three experiments, and are significant (p < 0.05 by log-rank with Bonferroni correction) in 2 out of 4 individual trials (Supplementary Table 2). (PNG 153 kb)

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Supplementary Table 1

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Supplementary Table 6

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Kruempel, J., Miller, H.A., Schaller, M.L. et al. Hypoxic response regulators RHY-1 and EGL-9/PHD promote longevity through a VHL-1-independent transcriptional response. GeroScience 42, 1621–1633 (2020). https://doi.org/10.1007/s11357-020-00194-0

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