Hypoxia-inducible factor induces cysteine dioxygenase and promotes cysteine homeostasis in Caenorhabditis elegans
- Pediatrics and Rare Diseases Group, Sanford Research, United States
- Department of Pediatrics, Sanford School of Medicine, University of South Dakota, United States
- Department of Molecular Biology, Massachusetts General Hospital, United States
Figures
Figure 1 with 1 supplement
egl-9 and rhy-1 inhibit cdo-1 transcription.
(A) Pathway for sulfur amino acid metabolism beginning with methionine. We highlight the roles of cystathionase (CTH-2/CTH), cysteine dioxygenase (CDO-1/CDO1), and the Moco-requiring sulfite oxidase …
Figure 1—figure supplement 1
The Pcdo-1::CDO-1::GFP transgene encodes a functional cysteine dioxygenase enzyme.
moc-1(ok366) cdo-1(mg622) double mutant animals expressing Pcdo-1::CDO-1::GFP or Pcdo-1::CDO-1[C85Y]::GFP transgenes were cultured from synchronized L1 larvae for 72 hr on wild-type (black, Moco+) …
Figure 2
cdo-1 transcription is activated by HIF-1 downstream of RHY-1, CYSL-1, and EGL-9.
(A) Expression of the Pcdo-1::GFP transgene is displayed for wild-type, egl-9(sa307), egl-9(sa307) hif-1(ia4) double mutant, egl-9(sa307); cysl-1(ok762) double mutant, rhy-1(ok1402), rhy-1(ok1402); …
Figure 3
High levels of cysteine activate cdo-1 transcription and are lethal to hif-1 and cysl-1 mutant animals.
(A) Expression of the Pcdo-1::GFP transgene is displayed for wild-type, egl-9(sa307), and rhy-1(ok1402) young-adult C. elegans exposed to 0 or 100 μM supplemental cysteine. Scale bar is 250 μm. …
Figure 4 with 2 supplements
Hypodermal CDO-1 accumulates in the cytoplasm when egl-9 or rhy-1 are inactive and is sufficient to promote sulfur amino acid metabolism.
(A) Diagram of cdo-1(rae273), a CRISPR/Cas9-generated allele with GFP inserted into the cdo-1 gene, creating a functional C-terminal CDO-1::GFP fusion protein expressed from the native cdo-1 locus. …
Figure 4—figure supplement 1
A functional CDO-1::GFP fusion protein is induced by loss of egl-9 or rhy-1.
(A) Expression of CDO-1::GFP from the cdo-1(rae273) allele is displayed for wild-type, egl-9(sa307), and rhy-1(ok1402) animals at the L4 stage of development. Scale bar is 250 μm. For GFP imaging, …
Figure 4—figure supplement 2
CDO-1::GFP encoded by cdo-1(rae273) is induced by supplemental cysteine.
(A) Expression of CDO-1::GFP from the cdo-1(rae273) allele is displayed for wild-type young adult animals exposed to 0 or 100 μM supplemental cysteine. Scale bar is 250 μm. For GFP imaging, exposure …
Figure 5
egl-9 inhibits cdo-1 transcription in a largely prolyl-hydroxylase and VHL-1-independent manner.
(A) The pathway of HIF-1 processing during normoxia is displayed. EGL-9 uses O2 as a substrate to hydroxylate (-OH) HIF-1 on specific proline residues. Prolyl hydroxylated HIF-1 is bound by VHL-1 …
Figure 6
Model for the regulation of cysteine metabolism by HIF-1.
(A) Proposed genetic pathway for the regulation of cdo-1. rhy-1, cysl-1, and egl-9 act in a negative-regulatory cascade to control activity of the HIF-1 transcription factor, which activates …
Tables
Table 1
Growth of C. elegans strains on standard laboratory conditions.
C. elegans strains and their corresponding mutations are displayed. For each strain, 5 L4-stage animals were seeded onto standard NGM petri dishes seeded with a monoculture of E. coli OP50. Petri …
| C. elegans strain | Genetic locus 1 | Genetic locus 2 | Genetic locus 3 | Days for population to starve petri dish ±SD (n) | Significant difference compared to GR2269 (Adjusted p Value) |
|---|---|---|---|---|---|
| N2 | Wild type | Wild type | Wild type | 5±1 (8) | No test |
| GR2269 | Wild type | suox-1(gk738847) | Wild type | 8±1 (4) | No test |
| JT307 | egl-9(sa307) | Wild type | Wild type | 7±1 (5) | No test |
| USD421 | egl-9(sa307) | suox-1(gk738847) | Wild type | 19±5 (11) | 0.0003 (***) |
| USD926 | egl-9(rae276) | Wild type | Wild type | 6±1 (3) | No test |
| USD937 | egl-9(rae276) | suox-1(gk738847) | Wild type | 9±1 (3) | 0.93 (ns) |
| USD512 | rhy-1(ok1402) | Wild type | Wild type | 6±0 (4) | No test |
| USD414 | rhy-1(ok1402) | suox-1(gk738847) | Wild type | 7±0 (4) | 0.99 (ns) |
| CB5602 | vhl-1(ok161) | Wild type | Wild type | 6±0 (4) | No test |
| USD422 | vhl-1(ok161) | suox-1(gk738847) | Wild type | 8±1 (4) | 0.99 (ns) |
| C. elegans strain | Genetic locus 1 | Genetic locus 2 | Genetic locus 3 | Days for population to starve petri dish ±SD (n) | Significant difference compared to USD421 (Adjusted p value) |
| USD421 | egl-9(sa307) | suox-1(gk738847) | Wild type | 19±5 (11) | No test |
| USD430 | egl-9(sa307) | suox-1(gk738847) | cdo-1(mg622) | 7±0 (6) | <0.0001 (****) |
| USD433 | egl-9(sa307) | suox-1(gk738847) | cth-2(mg599) | 9±1 (4) | <0.0001 (****) |
| USD432 | egl-9(sa307) | suox-1(gk738847) | rhy-1(ok1402) | 7±1 (4) | <0.0001 (****) |
| USD434 | egl-9(sa307) | suox-1(gk738847) | cysl-1(ok762) | 16±1 (9) | 0.1 (ns) |
| USD431 | egl-9(sa307) | suox-1(gk738847) | hif-1(ia4) | 9±1 (6) | <0.0001 (****) |
