Jumonji domain-containing protein 6 (Jmjd6) is required for angiogenic sprouting and regulates splicing of VEGF-receptor 1

Abstract

JmjC domain-containing proteins play a crucial role in the control of gene expression by acting as protein hydroxylases or demethylases, thereby controlling histone methylation or splicing. Here, we demonstrate that silencing of Jumonji domain-containing protein 6 (Jmjd6) impairs angiogenic functions of endothelial cells by changing the gene expression and modulating the splicing of the VEGF-receptor 1 (Flt1). Reduction of Jmjd6 expression altered splicing of Flt1 and increased the levels of the soluble form of Flt1, which binds to VEGF and placental growth factor (PlGF) and thereby inhibits angiogenesis. Saturating VEGF or PlGF or neutralizing antibodies directed against soluble Flt1 rescued the angiogenic defects induced by Jmjd6 silencing. Jmjd6 interacts with the splicing factors U2AF65 that binds to Flt1 mRNA. In conclusion, Jmjd6 regulates the splicing of Flt1, thereby controlling angiogenic sprouting.

Glutaredoxin-1 Up-regulation Induces Soluble Vascular Endothelial Growth Factor Receptor 1, Attenuating Post-ischemia Limb Revascularization

“…Soluble Flt production is regulated by alternative splicing (35), and ectodomain shedding of Flt by proteolytic cleavage , and VEGFR2 (D) in non-ischemic (NI) and ischemic (ISC) gastrocnemius muscles from WT and Glrx TG mice 14 days after HLI (n ϭ 6, p Ͻ 0.05). E and F, ELISA for sFLT1 (E) and VEGF (F) in plasma from WT and Glrx TG mice at 4 days (n ϭ 6) and 14 days (n ϭ 12) after HLI (*, p Ͻ 0.05).…”

“…Expression was obtained and analyzed using comparative C T (⌬⌬C T ) with StepOne TM real-time PCR software (Applied Biosystems), normalized to ␤-actin. The sequence for the custom designed TaqMan TM assay to assess human sFlt transcript waskindlyprovidedbyDr.StefanieDimmeler (35).SYBRgreenprimers were used to assess murine mFlt and sFlt according to the transcripts as published (24).…”

Vitamin D has wide regulatory effects on histone demethylase genes

“…Thus, De Santa et al 41 demonstrated that although neither Jmjd3 induction nor Jmjd3 depletion caused any detectable modification in the global levels of H3K27me3, Jmjd3 induction by LPS coincides with H3K27me3 downregulation and with Jmjd3 occupancy at the Bmp2 gene promoter. Similarly, Boeckel et al 61 did not observe changes in global histone methylation patterns in JMJD6-deficient cells, but they did not rule out the possibility that histone modifications might occur at specific promoters. Altogether, these results suggest that effects on H3K27 methylation must be analyzed at the single gene level.…”

“…[56][57][58][59] JMJD5 is required for breast cancer cell proliferation, while JMJD6 overexpression enhanced angiogenic sprouting in endothelial cells. 60,61 There is scarce literature for several of these genes, as the research on the biological role of this class of epigenetic Table S1. SDHA was used for normalization.…”

Clipping of arginine-methylated histone tails by JMJD5 and JMJD7

“…For example, JMJD6 has been reported variously to be an arginine demethylase (10,11), a hydroxylase (12,13), a single-stranded RNA-binding protein (14), or RNA demethylase (11). Although JMJD6 has been reported to possess histone arginine demethylase activity (10,11), we and others have been unable to replicate this result (12,(14)(15)(16)(17). Thus, the identities of proteins that reverse methylation of arginine residues, either through conventional demethylation or others, have not been resolved.…”