Lysyl 5-Hydroxylation, a Novel Histone Modification, by Jumonji Domain Containing 6 (JMJD6)*

Motoko Unoki,Akiko Masuda,Naoshi Dohmae,Kyohei Arita,Masanori Yoshimatsu,Yukiko Iwai,Yoshinori Fukui,Koji Ueda,Ryuji Hamamoto,Masahiro Shirakawa,Hiroyuki Sasaki,Yusuke Nakamura

doi:10.1074/jbc.m112.433284

Abstract

JMJD6 is reported to hydroxylate lysyl residues of a splicing factor, U2AF65. In this study, we found that JMJD6 hydroxylates histone lysyl residues. In vitro experiments showed that JMJD6 has a binding affinity to histone proteins and hydroxylates multiple lysyl residues of histone H3 and H4 tails. Using JMJD6 knock-out mouse embryos, we revealed that JMJD6 hydroxylates lysyl residues of histones H2A/H2B and H3/H4 in vivo by amino acid composition analysis. 5-Hydroxylysine was detected at the highest level in histones purified from murine testis, which expressed JMJD6 at a significantly high level among various tissues examined, and JMJD6 overexpression increased the amount of 5-hydroxylysine in histones in human embryonic kidney 293 cells. These results indicate that histones are additional substrates of JMJD6 in vivo. Because 5-hydroxylation of lysyl residues inhibited N-acetylation and N-methylation by an acetyltransferase and a methyltransferase, respectively, in vitro, histone 5-hydroxylation may have important roles in epigenetic regulation of gene transcription or chromosomal rearrangement.

Highlights

JMJD6 hydroxylates U2AF65, but its role in histone modification has been obscure
Using JMJD6 knock-out mouse embryos, we revealed that JMJD6 hydroxylates lysyl residues of histones H2A/H2B and H3/H4 in vivo by amino acid composition analysis. 5-Hydroxylysine was detected at the highest level in histones purified from murine testis, which expressed JMJD6 at a significantly high level among various tissues examined, and JMJD6 overexpression increased the amount of 5-hydroxylysine in histones in human embryonic kidney 293 cells
JMJD6 Effectively Hydroxylates Histone Lysyl Residues in Vitro—During screening of UHRF1-interacting proteins, we identified JMJD6 as a novel binding partner of UHRF1

Summary

Background

Results: Our analysis of histones purified from JMJD6 knock-out mouse embryos reveals that JMJD6 hydroxylates histone lysyl residues. Using JMJD6 knock-out mouse embryos, we revealed that JMJD6 hydroxylates lysyl residues of histones H2A/H2B and H3/H4 in vivo by amino acid composition analysis. Jumonji domain containing 6 (JMJD6), which possesses high binding affinity to single-stranded RNA, is reported to hydroxylate lysyl residues of an RNA splicing factor, U2AF65. Using JMJD6 knock-out mice, we revealed that JMJD6 hydroxylates histone lysyl residues and generates 5-hydroxylysine in vivo. Because it interfered with N-acetylation and N-methylation by an acetyltransferase and a methyltransferase, respectively, the modification may regulate transcription through these interactions with other histone modifications

EXPERIMENTAL PROCEDURES

RESULTS

DISCUSSION