| 2007 |
JMJD6 is an iron- and 2-oxoglutarate-dependent dioxygenase (JmjC family) that demethylates histone H3 at arginine 2 (H3R2) and histone H4 at arginine 3 (H4R3) in both biochemical and cell-based assays. |
In vitro biochemical demethylase assay and cell-based assays with purified JMJD6 |
Science |
High |
17947579
|
| 2009 |
JMJD6 catalyzes Fe(II)- and 2-oxoglutarate-dependent lysyl-5-hydroxylation of the splicing factor U2AF65, and this activity modulates alternative RNA splicing of specific endogenous and reporter genes. |
Mass spectrometry identification of hydroxylated lysine on U2AF65; in vitro hydroxylase assay; splicing reporter assays in cells with JMJD6 knockdown/overexpression |
Science |
High |
19574390
|
| 2010 |
Crystal structure of the JMJD6 catalytic domain in complex with Ni(II) (substituting Fe(II)) reveals a double-stranded beta-helical fold; mutational studies show how active-site geometry favors C-5 lysyl hydroxylation rather than N-epsilon demethylation. |
X-ray crystallography; active-site mutagenesis |
Journal of Molecular Biology |
High |
20684070 20685276
|
| 2010 |
JMJD6 binds efficiently to single-stranded RNA but not to single-stranded DNA, double-stranded RNA, or double-stranded DNA; the crystal structure reveals a novel substrate-binding groove and two positively charged surfaces with a stack of aromatic residues near the active center. |
X-ray crystallography; RNA/DNA binding assays; truncation analysis |
Proceedings of the National Academy of Sciences |
High |
20679243
|
| 2010 |
Jmjd6 is distributed throughout the nucleoplasm outside heterochromatic regions, localizes occasionally to nucleoli, is excluded from the nucleus during mitosis and reappears in telophase, and forms homo-multimers. Jmjd6 does NOT demethylate histone lysine residues H3K4, H3K9, H3K27, H3K36, or H4K20. |
Immunolocalization; Western blot; comparison of histone methylation states in wildtype vs. Jmjd6-knockout cells; overexpression of active and inactive forms |
PLoS One |
Medium |
21060799
|
| 2011 |
JMJD6-catalyzed hydroxylation of RNA-splicing regulatory protein fragments produces 5S-hydroxylysine, a stereochemistry distinct from collagen lysyl hydroxylases (which produce 5R products), indicating a distinct subfamily of lysyl hydroxylases. |
Amino acid analysis; NMR stereochemical assignment of hydroxylysine products from in vitro JMJD6 reactions |
ChemBioChem |
High |
22238144
|
| 2011 |
Jmjd6 silencing impairs angiogenic functions of endothelial cells by altering splicing of VEGF-receptor 1 (Flt1), increasing levels of soluble Flt1 that inhibits angiogenesis. Jmjd6 physically interacts with splicing factor U2AF65 that binds Flt1 mRNA. |
siRNA knockdown; RT-PCR splicing assay; co-immunoprecipitation of Jmjd6 with U2AF65; rescue experiments with VEGF/PlGF or anti-sFlt1 antibodies |
Proceedings of the National Academy of Sciences |
High |
21300889
|
| 2012 |
JMJD6 cannot demethylate histone arginine residues in vitro but does hydroxylate the histone H4 tail at lysine residues in a 2-OG- and Fe(II)-dependent manner. Homo-oligomerization of Jmjd6 requires its enzymatic (hydroxylase) activity and both N- and C-termini; Jmjd6 autodydroxylates its own N-terminus to form intermolecular covalent bonds mediating oligomerization. |
MALDI-TOF mass spectrometry in vitro enzymatic assay; mutational analysis of oligomerization domains |
Journal of Cellular Biochemistry |
Medium |
22189873
|
| 2013 |
JMJD6 hydroxylates multiple lysyl residues of histone H3, H4, H2A, and H2B tails in vitro and in vivo. 5-Hydroxylysine on histones inhibits N-acetylation and N-methylation by acetyltransferase and methyltransferase in vitro, suggesting cross-talk between histone modifications. |
In vitro hydroxylase assay; amino acid composition analysis of histones from JMJD6-knockout mouse embryos; JMJD6 overexpression in HEK293 cells; in vitro competition assays with acetyltransferase/methyltransferase |
Journal of Biological Chemistry |
High |
23303181
|
| 2013 |
JMJD6 and BRD4 co-occupy a unique cohort of distal enhancers (anti-pause enhancers, A-PEs) and regulate promoter-proximal Pol II pause release. BRD4-dependent JMJD6 recruitment to A-PEs mediates erasure of H4R3me2(s), which is read by 7SK snRNA; JMJD6 also decaps/demethylates 7SK snRNA, releasing the 7SK/HEXIM inhibitory complex and activating P-TEFb. |
ChIP-seq; RNA-seq; co-IP of JMJD6 with BRD4 and P-TEFb; 7SK snRNA demethylation/decapping assay; long-range chromatin interaction analysis |
Cell |
High |
24360279
|
| 2013 |
The polyserine (polyS) domain of Jmjd6 mediates its subnuclear localization; Jmjd6 lacking the polyS domain localizes to the nucleolus (fibrillar centre) and interacts with nucleolar proteins. Homo-oligomerization of Jmjd6 occurs in cells and the structure of oligomers changes without the polyS domain. |
Live-cell imaging; co-immunoprecipitation; F2H (fluorescent 2-hybrid) assay; deletion mutagenesis |
Biochemical Journal |
Medium |
23688307
|
| 2014 |
JMJD6 physically associates with p53 and acts as an alpha-ketoglutarate- and Fe(II)-dependent lysyl hydroxylase to hydroxylate p53 primarily at lysine 382. This hydroxylation antagonizes p53 acetylation, promotes p53 association with its negative regulator MDMX, and represses p53 transcriptional activity. |
Co-immunoprecipitation; in vitro hydroxylase assay; mass spectrometry identification of hydroxylation site; p53 acetylation assays; MDMX co-IP; reporter assays; JMJD6 knockdown/overexpression with phenotypic readouts |
PLoS Biology |
High |
24667498
|
| 2014 |
Upon estrogenic stimulation, JMJD6 interacts with methylated estrogen receptor alpha (ERα) and acts as an arginine demethylase to demethylate ERα, regulating rapid extranuclear estrogenic responses. |
Co-immunoprecipitation; JMJD6 silencing combined with in vitro demethylation assay |
PLoS One |
Medium |
24498420
|
| 2014 |
Jmjd6 interacts with multiple SR and SR-related proteins through their arginine-serine-rich (RS) domains, including U2AF65, Luc7L3, SRSF11, and Acinus S', but not the bona fide RS domain of SRSF1. Jmjd6 modifies constitutive splicing, binds RNA from a reporter plasmid, and co-localizes with nascent RNA. |
Co-immunoprecipitation; in vitro binding assay; splicing reporter assay; RNA binding assay; immunofluorescence co-localization with nascent RNA |
Nucleic Acids Research |
Medium |
24914048
|
| 2015 |
Jmjd6, acting as a lysyl hydroxylase for splicing regulatory proteins, is required for efficient splicing of intron 2 of the Aire gene in medullary thymic epithelial cells; Jmjd6 deficiency results in retention of Aire intron 2, markedly reduced mature Aire protein, and spontaneous multi-organ autoimmunity in mice. |
Jmjd6 conditional knockout mouse; RT-PCR splicing analysis; Western blot of Aire protein; phenotypic analysis of autoimmunity |
Nature Communications |
High |
26531897
|
| 2015 |
In Xenopus embryogenesis, Jmjd6 interacts with Tcf7l1 (Tcf3), competing with Groucho corepressor for the same binding region on Tcf7l1 to derepress Wnt target genes. Loss of Jmjd6 function causes anteroposterior body axis defects. |
Co-immunoprecipitation; reporter gene assays; morpholino loss-of-function in Xenopus embryos; rescue experiments |
Journal of Biological Chemistry |
Medium |
26157142
|
| 2017 |
JMJD6 is a novel stress granule (SG) component that interacts with G3BP1 complexes. JMJD6 promotes G3BP1 arginine demethylation (monomethylation and asymmetric dimethylation at three Arg residues) and thereby promotes SG formation; knockdown of JMJD6 represses SG formation, and rescue requires catalytically active but not mutant JMJD6. |
Co-immunoprecipitation; mass spectrometry; JMJD6 knockdown; rescue with catalytically active vs. inactive JMJD6; immunofluorescence of SG markers |
Journal of Biological Chemistry |
Medium |
28972166
|
| 2017 |
JMJD6 and BRD4 interact via the BRD4 extraterminal (ET) domain; NMR reveals a JMJD6 peptide (Lys84–Asn96) adopts an alpha-helix when bound to the ET domain through hydrophobic and electrostatic interactions. Single-stranded RNA binding by JMJD6 induces a conformational change that likely promotes this ET-domain interaction. |
NMR structure determination; mutagenesis; RNA binding assays |
Scientific Reports |
High |
29176719
|
| 2017 |
JMJD6 forms protein complexes with N-Myc and BRD4 in neuroblastoma cells and is important for transcription of E2F2, N-Myc, and c-Myc. JMJD6 gene is associated with transcriptional super-enhancers. |
Co-immunoprecipitation; ChIP-seq; gene knockdown with transcriptional readouts |
Nature Communications |
Medium |
31346162
|
| 2017 |
JMJD6 regulates the alternative splicing of PAK1, a MAPK pathway component, in melanoma cells; knockdown affects a panel of alternative splicing events as measured by RNA-seq. |
RNA-seq splicing analysis; JMJD6 knockdown; functional assays in melanoma cell lines |
Molecular Cancer |
Medium |
29187213
|
| 2017 |
JMJD6 co-regulates alternative splicing events together with U2AF65; JMJD6 enzymatic activity (lysine hydroxylation of U2AF65) is required for a subset of co-regulated splicing events but not all, indicating both enzymatic activity-dependent and -independent mechanisms of splicing regulation. Validated in jmjd6 knockout mice. |
RASL-Seq splicing profiling; JMJD6/U2AF65 knockdown; JMJD6 knockout mice; co-IP; mutagenesis of catalytic residues |
Nucleic Acids Research |
High |
27899633
|
| 2018 |
JMJD6 is required for ERα-bound active enhancer activation and RNA Pol II recruitment: JMJD6 interacts with MED12 in the mediator complex, is necessary for MED12 interaction with CARM1, and CARM1 methylates MED12 at multiple arginine sites to regulate chromatin binding, thereby enabling transcriptional pause release of estrogen target genes. |
Co-immunoprecipitation; ChIP-seq; enhancer RNA measurement; JMJD6 knockdown; CARM1 methylation assays; cell proliferation and xenograft assays |
Molecular Cell |
High |
29628309
|
| 2018 |
JMJD6 has intrinsic tyrosine kinase activity and phosphorylates histone H2A.X at tyrosine 39 (H2A.XY39ph), using ATP and GTP as phosphate donors; high JMJD6 promotes autophagy in triple-negative breast cancer cells via regulation of autophagy-related gene expression through the H2A.XY39ph axis. |
In vitro kinase assay; mass spectrometry identification of phosphorylation site; JMJD6 knockdown with autophagy readouts; cell growth assays |
Oncogene |
Medium |
30185813
|
| 2019 |
Biochemical (MS- and NMR-based kinetic assays) and structural (crystallographic) analyses with purified JMJD6 support assignment as a lysyl hydroxylase; the study did NOT observe N-methyl arginyl N-demethylation activity with purified JMJD6, calling into question its arginine demethylase activity. |
MS-based kinetic assay; NMR assay; X-ray crystallography; substrate fragment screening |
Journal of Biological Chemistry |
High |
31147442
|
| 2019 |
JMJD6 homo-oligomerization does not require catalytic activity; the AT hook-like domain (mediating DNA/RNA interaction) is required for JMJD6 chromatin binding and function in adipogenic differentiation, whereas catalytic activity and the polyserine and sumoylation domains are dispensable. |
Domain mutagenesis; oligomerization assays; chromatin immunoprecipitation; adipogenic differentiation assays in JMJD6 mutant-rescued cells |
PLoS One |
Medium |
31430278
|
| 2020 |
JMJD6 is recruited to DNA double-strand breaks (DSBs), controls spreading of histone ubiquitination and accumulation of repair proteins around DSBs, and promotes transcriptional silencing. Independently of catalytic activity, JMJD6 interacts with SIRT1 and recruits it to chromatin to downregulate H4K16ac around DSBs, modulating NHEJ and HR efficiency. |
Microirradiation with live imaging; co-immunoprecipitation of JMJD6 with SIRT1; ChIP for H4K16ac; NHEJ/HR reporter assays; catalytic mutant rescue |
Cell Death and Differentiation |
High |
31358914
|
| 2020 |
JMJD6 cleaves methylphosphate capping enzyme (MePCE), a core component of the 7SK snRNP complex, via a novel proteolytic activity. This cleavage releases P-TEFb from the 7SK snRNP complex; downstream effect is phosphorylation of Pol II CTD. Crystal structure of JMJD6 bound to methyl-arginine was determined. |
Crystal structure of JMJD6 bound to methyl-arginine; in vitro and in vivo MePCE cleavage assays; binding assays; Jmjd6 KO and overexpression with Pol II CTD phosphorylation readout |
eLife |
High |
32048991
|
| 2020 |
JMJD6 is rapidly recruited to nucleolar DNA damage sites and is required for relocation of rDNA into nucleolar caps, rDNA stability, and maintaining rDNA repeat integrity after irradiation. JMJD6 interacts with nucleolar protein Treacle and modulates its interaction with NBS1. |
Live-cell imaging after micro-irradiation; mass spectrometry interactome; JMJD6-deficient cells; rDNA repeat analysis by Southern blot/FISH |
PLoS Genetics |
Medium |
32598339
|
| 2021 |
JMJD6 knockdown reduces recruitment of U2AF65 to AR pre-mRNA and reduces AR-V7 splice variant levels; mutagenesis indicates JMJD6 catalytic activity is required for AR-V7 generation. The JMJD6/U2AF65 axis thus controls AR-V7 splicing in prostate cancer. |
siRNA knockdown; mutagenesis of catalytic residues; RIP (RNA immunoprecipitation) of U2AF65 on AR pre-mRNA; RT-PCR for AR-V7; prostate cancer cell growth assays |
Cancer Research |
Medium |
33822745
|
| 2021 |
JMJD6 negatively regulates antiviral innate immune signaling by recruiting the ubiquitin E3 ligase RNF5 to promote K48-linked ubiquitination and degradation of activated IRF3, thereby reducing type-I interferon production. |
Co-immunoprecipitation; ubiquitination assay; proteomic screen; JMJD6 KO via piggyBac transposon in mice; viral replication assays |
PLoS Pathogens |
Medium |
33684176
|
| 2021 |
JMJD6 is essential for short- and long-term maintenance of the hematopoietic stem cell (HSC) pool; Jmjd6-deficient HSCs fail to expand after injury and lose self-renewal upon serial transplantation. JMJD6 represses mitochondrial OXPHOS, protein synthesis, p53 stabilization, and mTORC1 signaling in HSCs; ROS elevation from OXPHOS causally mediates HSC failure. |
Hematopoietic-specific conditional Jmjd6 KO mouse; serial transplantation; metabolic assays (mitochondrial respiration, ROS); N-acetyl-L-cysteine rescue |
Blood Advances |
High |
33560400
|
| 2022 |
JMJD6 catalyzes lysine hydroxylation at 150 sites on 48 protein substrates identified by mass spectrometry, predominantly within unstructured lysine-rich regions. BRD4 is hydroxylated on 19 lysine sites. Nearly all identified substrates are associated with membraneless organelle formation, suggesting JMJD6 may regulate liquid-liquid phase separation. |
Mass spectrometry with lysine propionyl derivatization; nontryptic proteolysis; JMJD6-depleted vs. control cell comparison |
Proceedings of the National Academy of Sciences |
High |
35930668
|
| 2022 |
JMJD6 interacts with RBM39 and co-occupies the DGAT1 gene promoter with H3K4me3 to induce DGAT1 expression, thereby promoting lipid droplet formation and ccRCC tumorigenesis. |
siRNA screen; ChIP-seq; RNA-seq; co-immunoprecipitation of JMJD6 with RBM39; JMJD6 KD with lipid droplet and tumor growth readouts |
Molecular Cell |
Medium |
35764091
|
| 2022 |
JMJD6 demethylates HURP at R122, promoting Golgi apparatus repositioning (GR) and directional cell migration via a NF-κB-induced centrosome repositioning and subsequent Cdc42 upregulation cascade; PRMT5 methylation of HURP R122 opposes this. |
Co-immunoprecipitation; HURP methylation assays; JMJD6 KD; Golgi repositioning assays; cell migration assays; mechanistic epistasis using methylation-mimic/deficiency mutants |
Journal of Cellular Physiology |
Medium |
36250981
|
| 2022 |
JMJD6 regulates splicing of its own pre-mRNA to produce isoforms (JMJD6-2 and JMJD6-Ex5) with distinct C-terminal sequences and different interaction partners; JMJD6-2 interacts with SR-like splicing factors (RS-domain proteins) while JMJD6-Ex5 interacts with SMN complex, hnRNPs, UBF, and FCP1. JMJD6-2 but not JMJD6-Ex5 inhibits exon inclusion in a splicing reporter. |
Immunoprecipitation followed by LC-MS/MS; HIV-based splicing reporter assay; siRNA knockdown; RT-PCR isoform analysis |
International Journal of Molecular Sciences |
Medium |
32927736
|
| 2023 |
JMJD6 interacts with NF-κB p65 in the cytoplasm and demethylates p65 at arginine 149 (R149), inhibiting nuclear translocation of p65 and thereby suppressing NF-κB signaling and protecting against pathological cardiac hypertrophy. |
Co-immunoprecipitation of JMJD6 with p65; in vitro/in vivo demethylation assays; cardiac-specific JMJD6 overexpression in rats; sgRNA-mediated JMJD6 depletion; echocardiography; hypertrophic gene expression assays |
Acta Pharmacologica Sinica |
Medium |
37186122
|
| 2024 |
JMJD6 acts as a hub connecting pre-mRNA splicing and glutamine metabolism in MYC-driven neuroblastoma: it physically interacts with RNA-binding proteins involved in splicing and protein homeostasis, and controls alternative splicing of glutaminase (GLS) isoforms KGA and GAC (rate-limiting enzymes of glutaminolysis). JMJD6 also complexes with RBM39, linking it to sensitivity to the splicing inhibitor indisulam. |
Co-immunoprecipitation; RNA-seq/splicing analysis; metabolic assays; cell transformation assays; indisulam treatment with JMJD6-dependent readout |
eLife |
Medium |
38488852
|
| 2024 |
HSF1 directly binds JMJD6 gene and promotes its transcription; JMJD6 in turn reduces HSP70 R469 monomethylation to disrupt HSP70-HSF1 repressive complexes, enabling enhanced HSF1 activation in a positive feedback circuit mediating cellular adaptation to proteotoxic stress. |
Genome-wide RNAi screen with HSR reporter; ChIP assay for HSF1 at JMJD6 locus; JMJD6-mediated HSP70 demethylation assay; co-immunoprecipitation |
Proceedings of the National Academy of Sciences |
Medium |
38985769
|
| 2025 |
SPOP mutants impair proteasomal degradation of JMJD6; elevated JMJD6 and ATF4 coordinate enhancer-promoter loop interactions to stimulate glutathione biosynthesis pathway genes (SLC7A11, GCLM, ME1). JMJD6 recruits mediator subunits (Med1/Med14) to assemble de novo enhancers at these loci, conferring ferroptosis resistance independently of androgen receptor. |
Co-immunoprecipitation; ChIP-seq; enhancer-promoter loop analysis; JMJD6 depletion; preclinical tumor models with ferroptosis inducers |
Cancer Research |
Medium |
39903850
|
| 2025 |
JMJD6 is acetylated at lysine 375 by the acetyltransferase PCAF; K375 acetylation weakens JMJD6 arginine demethylase activity, enhances METTL14 expression, increases m6A modification of SLC3A2, and thereby promotes ferroptosis sensitivity in lung cancer cells. |
Co-immunoprecipitation; GST pulldown; in vitro acetylation assay; ChIP; MeRIP (m6A sequencing); functional ferroptosis assays; JMJD6 overexpression/knockdown |
Journal of Translational Medicine |
Medium |
40011892
|
| 2017 |
JMJD6 suppresses Myc-induced apoptosis through inhibition of p19ARF mRNA and protein, leading to reduced p53 levels; JMJD6 binds the p19ARF promoter and exerts its inhibitory function through demethylation of H4R3me2a at this promoter. |
Chromatin immunoprecipitation at p19ARF promoter; JMJD6 overexpression/knockdown with p19ARF/p53 readouts; cell viability assays under stress; xenograft tumor assays |
Clinical Epigenetics |
Medium |
27081402
|
| 2009 |
Endogenous Jmjd6 is expressed at the cell surface of immature monocyte-like THP-1 cells activated with PMA and translocates to the nucleus upon macrophage differentiation; antibody blockade of surface Jmjd6 suppresses phagocytosis of dead cells in immature macrophages. |
Immunocytochemistry; antibody blocking of endogenous surface Jmjd6; phagocytosis assay; time-course subcellular fractionation |
Journal of Cellular Physiology |
Medium |
19492415
|
| 2017 |
JMJD6 demethylates STAT1 arginine methylation, suppressing IFNα-induced interferon-stimulated gene (ISG) activation; overexpression of JMJD6 suppressed STAT1 methylation and ISG activation, while JMJD6 silencing enhanced both. |
JMJD6 overexpression/knockdown; STAT1 methylation assay; ISG expression measurement; HCV RNA quantification; in vivo mouse model |
Cellular and Molecular Gastroenterology and Hepatology |
Medium |
29693039
|
| 2017 |
JMJD6 binds to the HOTAIR promoter region (−123 to −103 bp) and induces HOTAIR transcription; a catalytically dead JMJD6 mutant (H187A) can bind the HOTAIR promoter but fails to activate transcription, demonstrating enzymatic activity-dependent transcriptional regulation. |
ChIP; EMSA; deletion constructs of HOTAIR promoter; luciferase reporter; JMJD6 overexpression/siRNA in multiple cell lines; catalytic mutant analysis |
Biochemical Journal |
Medium |
29229759
|
| 2020 |
Fibronectin is a novel substrate of JMJD6-mediated lysyl hydroxylation; JMJD6-catalyzed hydroxylation of fibronectin precedes its glycosylation, deposition, and degradation in placental mesenchymal cells. |
MALDI-TOF mass spectrometry identification of fibronectin as JMJD6 substrate; JMJD6 knockdown recapitulating fibronectin accumulation phenotype; iron rescue (Hinokitiol) experiments |
Frontiers in Cell and Developmental Biology |
Medium |
34055782
|