liu.seSearch for publications in DiVA
Change search
ReferencesLink to record
Permanent link

Direct link
Histone H5chromatin interactions in situ are strongly modulated by H5 C-terminal phosphorylation
Linköping University, Department of Clinical and Experimental Medicine. Linköping University, Faculty of Health Sciences.
Bulgarian Academic Science, Bulgaria .
Bulgarian Academic Science, Bulgaria .
Medical University of Innsbruck, Austria .
Show others and affiliations
2013 (English)In: Cytometry Part A, ISSN 1552-4922, Vol. 83A, no 3, 273-279 p.Article in journal (Refereed) Published
Abstract [en]

We used linker histone-depleted normal human fibroblast nuclei as templates to study how phosphorylation affects histone H5 binding to chromatin in situ. Permeabilized cells were treated with 0.7 M NaCl to extract the native linker histones. Histone H5 was purified from chicken erythrocytes and phosphorylated in vitro by recombinant cdk5/p35 kinase. High performance capillary electrophoresis (HPCE) showed that the phosphorylated protein contained a mixture of multiply phosphorylated forms. Control experiments, using mass spectrometry, revealed that up to five SPXK motifs in the C terminus were phosphorylated, but also that about 10% of the protein contained one phosphoserine in the N-terminus. Reconstitution of H1-depleted fibroblast nuclei with nonphosphorylated or phosphorylated H5 was performed at physiological ionic strength. The bound H5 was then extracted using NaCl concentrations in the range of 0.15 to 0.7 M. The release of the H5 molecules was monitored by DAPI staining and image cytofluorometry. Our results show that H5 phosphorylation substantially reduced its affinity for chromatin in situ, which support previous observations indicating that C-terminal phosphorylation may be essential for the biological functions of linker histones.

Place, publisher, year, edition, pages
Wiley-Blackwell , 2013. Vol. 83A, no 3, 273-279 p.
Keyword [en]
chromatin, linker histones, affinity, phosphorylation
National Category
Medical and Health Sciences
URN: urn:nbn:se:liu:diva-90191DOI: 10.1002/cyto.a.22221ISI: 000315299000004OAI: diva2:613158

Funding Agencies|Bulgarian National Science Fund|K-906/1999|Swedish Research Council|349-2001-6688|European Science Foundation EUROCORES Programme EuroDYNA (Austrian Science Foundation)|I23-B03|EC Sixth Framework Programme|ERAS-CT-2003-980409|

Available from: 2013-03-26 Created: 2013-03-21 Last updated: 2013-03-26

Open Access in DiVA

No full text

Other links

Publisher's full text

Search in DiVA

By author/editor
Kostova-Koleva, Nora N.Rundquist, Ingemar
By organisation
Department of Clinical and Experimental MedicineFaculty of Health SciencesCell Biology
In the same journal
Cytometry Part A
Medical and Health Sciences

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 41 hits
ReferencesLink to record
Permanent link

Direct link