OGFOD1

OGFOD1 is a Fe(II)- and 2-oxoglutarate-dependent prolyl hydroxylase that controls protein synthesis by post-translationally modifying the translation machinery and tuning ribosomal fidelity (PMID:24550447, PMID:40961937). It catalyzes trans-3 prolyl hydroxylation of Pro-62 in the small ribosomal subunit protein RPS23 (uS12), retaining high affinity for the hydroxylated product; loss of this catalytic activity triggers stress granule induction, translational arrest, and growth impairment, and the phenotype is rescued only by catalytically active enzyme (PMID:24550447). Crystal structures of the human enzyme with 2OG-oxygenase inhibitors define a prolyl-3-hydroxylase active site distinct from prolyl-4-hydroxylases (PMID:25728928). Beyond ribosome modification, OGFOD1 localizes to stress granules and associates with the ribosome and stress granule proteins, and interacts with eIF2α and its kinase HRI to set the level of eIF2α phosphorylation and the pace of polyribosome recovery after stress (PMID:20154146). It also directly binds the RNA Pol II C-terminal domain and is phosphorylated at Ser-256 by CDK7/CDK9, which is required for its enhancement of transcriptional activation and tumor growth (PMID:34298635). Functionally, OGFOD1 loss reshapes the proteome largely at the post-transcriptional level, controlling translation and stability of specific targets including cell-cycle regulators (CDK1 via HuR, with CDK2/CCNB1 reduced transcriptionally) and tissue-specific factors (PMID:31112528, PMID:36464654), and confers protection against ischemia-reperfusion injury through UPR/autophagy activation and altered β-alanine metabolism (PMID:34668514, PMID:38454480), and supports global protein synthesis in chemoresistant AML (PMID:40961937). Studies of the yeast ortholog Tpa1 establish a conserved double-stranded β-helix dioxygenase architecture coupled to translation termination and oxygen/hypoxia-responsive gene regulation (PMID:20040577, PMID:20630870).

1. 2009 Medium

   Defining the enzymatic scaffold: it was unknown how this 2OG-oxygenase family member was organized, and structures of the yeast ortholog Tpa1 revealed a bilobed double-stranded β-helix fold with cofactor binding restricted to the N-terminal domain, plus poly(rA) binding hinting at mRNA association.

   Evidence X-ray crystallography of S. cerevisiae Tpa1 with Fe(III) and 2OG, poly(rA) binding assay

   PMID:20040577

   Open questions at the time

   • Catalytic substrate not identified in this work
   • Ortholog study; human enzyme not addressed
   • Functional role of the inactive C-terminal domain unresolved
2. 2010 Medium

   Linking catalysis to translation and oxygen sensing: Tpa1 active-site integrity and its partner Ett1/Nro1 were shown to be essential for correct translation termination and for repression of Hap1-regulated hypoxic genes, connecting the dioxygenase to translation and oxygen-responsive transcription.

   Evidence Crystallography, active-site mutagenesis, genetic deletion with translation termination and gene-expression readouts in yeast

   PMID:20630870

   Open questions at the time

   • Direct catalytic substrate still unidentified
   • Mechanism connecting termination to oxygen sensing unclear
   • Human relevance inferred from ortholog
3. 2010 Medium

   Placing the human protein in the stress-response network: OGFOD1 was found in stress granules and bound to the ribosome, eIF2α, and HRI, with its level controlling eIF2α phosphorylation and translational recovery—establishing a role in the integrated stress response.

   Evidence Reciprocal Co-IP, siRNA knockdown and overexpression, polyribosome sedimentation, immunofluorescence

   PMID:20154146

   Open questions at the time

   • Whether eIF2α/HRI effects require catalytic activity not resolved
   • Single lab
   • Direct vs. indirect nature of HRI interaction not dissected
4. 2010 Medium

   Tying catalysis to a downstream transcript and an ischemic phenotype: OGFOD1 catalytic activity was shown to be required for ATPAF1 expression and ischemic cell death, with catalytic-mutant and ATPAF1 rescue experiments establishing causality.

   Evidence Gene silencing, cDNA microarray, wild-type vs. catalytic-mutant reintroduction, ATPAF1 rescue

   PMID:20579638

   Open questions at the time

   • Direct substrate connecting catalysis to ATPAF1 unknown
   • Mechanism of transcript-specific regulation unclear
   • Single lab
5. 2014 High

   Identifying the physiological substrate: it was unknown what OGFOD1 hydroxylates, and in vitro assays with MS established RPS23 (uS12) Pro-62 as the trans-3 prolyl hydroxylation target, with catalytic-mutant rescue tying the modification to stress granule and growth phenotypes.

   Evidence In vitro hydroxylation assay, mass spectrometry, Co-IP, knockdown/rescue with catalytic mutants

   PMID:24550447

   Open questions at the time

   • How Pro-62 hydroxylation alters ribosome function mechanistically not fully defined
   • Whether other substrates exist not excluded
6. 2014 Medium

   Establishing a non-ribosomal catalytic capacity in the ortholog: Tpa1 was shown to directly repair methylated DNA in vitro in a cofactor-dependent manner and to act in a pathway parallel to base excision repair.

   Evidence In vitro DNA repair assay with purified Tpa1, active-site mutagenesis, genetic epistasis

   PMID:25381260

   Open questions at the time

   • Whether human OGFOD1 has analogous DNA-repair activity untested
   • Physiological significance versus RPS23 hydroxylation unclear
   • Ortholog study
7. 2015 High

   Defining the structural basis of substrate specificity: human OGFOD1 structures with broad-spectrum 2OG-oxygenase inhibitors revealed the prolyl-3 hydroxylase active site and its distinctions from prolyl-4 hydroxylases, enabling selective inhibitor design.

   Evidence X-ray crystallography of human OGFOD1 with NOG and 2,4-PDCA, structural comparison

   PMID:25728928

   Open questions at the time

   • No structure with RPS23 substrate bound
   • Selective inhibitors not yet developed in this work
8. 2019 Medium

   Demonstrating proteome-wide post-transcriptional control: OGFOD1 deletion in cardiomyocytes selectively altered translation and splicing with poor mRNA-protein correlation, establishing post-transcriptional regulation as its primary cellular consequence and a role in cardiac proteome composition.

   Evidence OGFOD1 deletion in iPSC-cardiomyocytes, RNA-seq, quantitative proteomics, correlation analysis

   PMID:31112528

   Open questions at the time

   • Which targets are direct consequences of RPS23 hydroxylation unclear
   • Mechanism of selective translational control undefined
   • Single lab
9. 2021 Medium

   Uncovering a transcriptional arm: OGFOD1 was shown to bind the RNA Pol II CTD and to be phosphorylated by CDK7/CDK9 at Ser-256, with a non-phosphorylatable mutant failing to enhance transcription and tumor growth.

   Evidence Co-IP, CDK7/CDK9 phosphorylation assay, S256A phospho-mutant rescue, tumor growth assays

   PMID:34298635

   Open questions at the time

   • Relationship between CTD binding and prolyl hydroxylase activity unresolved
   • Whether OGFOD1 hydroxylates a transcriptional target unknown
   • Single lab
10. 2022 Medium

    Connecting OGFOD1 to cell-cycle control and cancer: knockdown induced arrest through post-transcriptional CDK1 loss (via HuR) and transcriptional CDK2/CCNB1 depletion with p21 nuclear accumulation, dissecting dual regulatory modes.

    Evidence siRNA knockdown, mRNA stability assays, RT-qPCR, p21 immunofluorescence, RNP-IP with HuR

    PMID:36464654

    Open questions at the time

    • Direct molecular link between OGFOD1 catalysis and HuR-mediated regulation unknown
    • Single cancer model
    • Single lab
11. 2022 Medium

    Defining a metabolic basis for cardioprotection: OGFOD1 knockout mice showed β-alanine accumulation, reduced infarct size, and improved function after ischemia-reperfusion, with carnosine rescue implicating β-alanine metabolism in the protection.

    Evidence Knockout mouse, TMT proteomics, metabolomics, ex vivo/in vivo ischemia-reperfusion, carnosine pharmacological rescue

    PMID:34668514

    Open questions at the time

    • How OGFOD1 catalysis controls β-alanine metabolism unclear
    • Link between RPS23 hydroxylation and metabolic phenotype undefined
    • Single lab
12. 2024 Medium

    Mapping the protective signaling output and a druggable target: OGFOD1 knockdown protected against ischemic stroke by activating UPR and autophagy independently of HIF-1α, and FG4592 was identified as an OGFOD1 inhibitor.

    Evidence Molecular docking, OGFOD1 knockdown in vitro/in vivo, mouse ischemic stroke model, UPR/autophagy blockade, HIF-1α knockdown

    PMID:38454480

    Open questions at the time

    • FG4592 binding to OGFOD1 is computational/pharmacological, not structurally confirmed
    • Mechanism linking inhibition to UPR activation unclear
    • Single lab
13. 2025 Medium

    Establishing therapeutic relevance in leukemia: OGFOD1 was shown to drive global protein synthesis in AML through RPS23 Pro-62 hydroxylation, and its inhibition impaired translation and improved survival in chemoresistant models while sparing normal hematopoiesis.

    Evidence Patient-derived xenografts, genetic inhibition, proteomics, translation assays

    PMID:40961937

    Open questions at the time

    • Therapeutic window in patients untested
    • Selective inhibitor not yet available
    • Single study
14. 2026 Medium

    Extending post-transcriptional control to hepatic ischemia: OGFOD1 silencing reduced SPARC protein without changing its mRNA, and SPARC overexpression rescued apoptosis and oxidative stress, placing SPARC downstream in a post-transcriptional pathway.

    Evidence siRNA silencing in H/R hepatocytes and HIRI rat model, combined transcriptomics/proteomics, SPARC overexpression rescue

    PMID:41558616

    Open questions at the time

    • Mechanism by which OGFOD1 controls SPARC translation/stability unknown
    • Whether RPS23 hydroxylation mediates this effect untested
    • Single lab

• It remains unresolved how RPS23 Pro-62 hydroxylation mechanistically connects the enzyme's many downstream phenotypes—selective translation, transcriptional enhancement via Pol II CTD, metabolic and UPR/autophagy outputs—and whether the transcriptional and DNA-related activities require its catalytic function.
• No structure of OGFOD1 bound to RPS23
• Catalytic dependence of CTD-binding and metabolic phenotypes not established
• Direct mechanism linking ribosomal modification to transcript-specific regulation unknown