2.50
Hdl Handle:
http://hdl.handle.net/10033/117145
Title:
Analysis of Jmjd6 cellular localization and testing for its involvement in histone demethylation.
Authors:
Hahn, Phillip; Wegener, Ivonne; Burrells, Alison; Böse, Jens; Wolf, Alexander; Erck, Christian; Butler, Danica; Schofield, Christopher J; Böttger, Angelika; Lengeling, Andreas
Abstract:
BACKGROUND: Methylation of residues in histone tails is part of a network that regulates gene expression. JmjC domain containing proteins catalyze the oxidative removal of methyl groups on histone lysine residues. Here, we report studies to test the involvement of Jumonji domain-containing protein 6 (Jmjd6) in histone lysine demethylation. Jmjd6 has recently been shown to hydroxylate RNA splicing factors and is known to be essential for the differentiation of multiple tissues and cells during embryogenesis. However, there have been conflicting reports as to whether Jmjd6 is a histone-modifying enzyme. METHODOLOGY/PRINCIPAL FINDINGS: Immunolocalization studies reveal that Jmjd6 is distributed throughout the nucleoplasm outside of regions containing heterochromatic DNA, with occasional localization in nucleoli. During mitosis, Jmjd6 is excluded from the nucleus and reappears in the telophase of the cell cycle. Western blot analyses confirmed that Jmjd6 forms homo-multimers of different molecular weights in the nucleus and cytoplasm. A comparison of mono-, di-, and tri-methylation states of H3K4, H3K9, H3K27, H3K36, and H4K20 histone residues in wildtype and Jmjd6-knockout cells indicate that Jmjd6 is not involved in the demethylation of these histone lysine residues. This is further supported by overexpression of enzymatically active and inactive forms of Jmjd6 and subsequent analysis of histone methylation patterns by immunocytochemistry and western blot analysis. Finally, treatment of cells with RNase A and DNase I indicate that Jmjd6 may preferentially associate with RNA/RNA complexes and less likely with chromatin. CONCLUSIONS/SIGNIFICANCE: Taken together, our results provide further evidence that Jmjd6 is unlikely to be involved in histone lysine demethylation. We confirmed that Jmjd6 forms multimers and showed that nuclear localization of the protein involves association with a nucleic acid matrix.
Affiliation:
Department of Experimental Mouse Genetics, Helmholtz Centre for Infection Research, Braunschweig, Germany.
Citation:
Analysis of Jmjd6 cellular localization and testing for its involvement in histone demethylation. 2010, 5 (10):e13769 PLoS ONE
Journal:
PloS one
Issue Date:
2010
URI:
http://hdl.handle.net/10033/117145
DOI:
10.1371/journal.pone.0013769
PubMed ID:
21060799
Type:
Article
Language:
en
ISSN:
1932-6203
Appears in Collections:
publications of the department infection genetics (INFG)

Full metadata record

DC FieldValue Language
dc.contributor.authorHahn, Phillipen
dc.contributor.authorWegener, Ivonneen
dc.contributor.authorBurrells, Alisonen
dc.contributor.authorBöse, Jensen
dc.contributor.authorWolf, Alexanderen
dc.contributor.authorErck, Christianen
dc.contributor.authorButler, Danicaen
dc.contributor.authorSchofield, Christopher Jen
dc.contributor.authorBöttger, Angelikaen
dc.contributor.authorLengeling, Andreasen
dc.date.accessioned2010-12-03T15:03:16Z-
dc.date.available2010-12-03T15:03:16Z-
dc.date.issued2010-
dc.identifier.citationAnalysis of Jmjd6 cellular localization and testing for its involvement in histone demethylation. 2010, 5 (10):e13769 PLoS ONEen
dc.identifier.issn1932-6203-
dc.identifier.pmid21060799-
dc.identifier.doi10.1371/journal.pone.0013769-
dc.identifier.urihttp://hdl.handle.net/10033/117145-
dc.description.abstractBACKGROUND: Methylation of residues in histone tails is part of a network that regulates gene expression. JmjC domain containing proteins catalyze the oxidative removal of methyl groups on histone lysine residues. Here, we report studies to test the involvement of Jumonji domain-containing protein 6 (Jmjd6) in histone lysine demethylation. Jmjd6 has recently been shown to hydroxylate RNA splicing factors and is known to be essential for the differentiation of multiple tissues and cells during embryogenesis. However, there have been conflicting reports as to whether Jmjd6 is a histone-modifying enzyme. METHODOLOGY/PRINCIPAL FINDINGS: Immunolocalization studies reveal that Jmjd6 is distributed throughout the nucleoplasm outside of regions containing heterochromatic DNA, with occasional localization in nucleoli. During mitosis, Jmjd6 is excluded from the nucleus and reappears in the telophase of the cell cycle. Western blot analyses confirmed that Jmjd6 forms homo-multimers of different molecular weights in the nucleus and cytoplasm. A comparison of mono-, di-, and tri-methylation states of H3K4, H3K9, H3K27, H3K36, and H4K20 histone residues in wildtype and Jmjd6-knockout cells indicate that Jmjd6 is not involved in the demethylation of these histone lysine residues. This is further supported by overexpression of enzymatically active and inactive forms of Jmjd6 and subsequent analysis of histone methylation patterns by immunocytochemistry and western blot analysis. Finally, treatment of cells with RNase A and DNase I indicate that Jmjd6 may preferentially associate with RNA/RNA complexes and less likely with chromatin. CONCLUSIONS/SIGNIFICANCE: Taken together, our results provide further evidence that Jmjd6 is unlikely to be involved in histone lysine demethylation. We confirmed that Jmjd6 forms multimers and showed that nuclear localization of the protein involves association with a nucleic acid matrix.en
dc.language.isoenen
dc.titleAnalysis of Jmjd6 cellular localization and testing for its involvement in histone demethylation.en
dc.typeArticleen
dc.contributor.departmentDepartment of Experimental Mouse Genetics, Helmholtz Centre for Infection Research, Braunschweig, Germany.en
dc.identifier.journalPloS oneen

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