In vitro binding of H1 histone subtypes to nucleosomal organized mouse mammary tumor virus long terminal repeat promotor.
The binding of all known linker histones, named H1a through H1e, including H1(0) and H1t, to a model chromatin complex based on a DNA fragment containing the mouse mammary tumor virus long terminal repeat promotor was systematically studied. As for the histone subtype H1b, we found a dissociation constant of 8-16 nM to a single mononucleosome (210 base pairs), whereas the binding constant of all other subtypes varied between 2 and 4 nM. Most of the H1 histones, namely H1a, H1c, H1d/e, and H1(0), completely aggregate polynucleosomes (1.3 kilobase pairs, 6 nucleosomes) at 270-360 nM, corresponding to a molar ratio of six to eight H1 molecules per reconstituted nucleosome. To form aggregates with the histones H1t and H1b, however, greater amounts of protein were required. Furthermore, our results show that specific types of in vivo phosphorylation of the linker histone tails influence both the binding to mononucleosomes and the aggregation of polynucleosomes. S phase-specific phosphorylation with one to three phosphate groups at specific sites in the C terminus influences neither the binding to a mononucleosome nor the aggregation of polynucleosomes. In contrast, highly phosphorylated H1 histones with four to five phosphate groups in the C and N termini reveal a very high binding affinity to a mononucleosome but a low chromatin aggregation capability. These findings suggest that specific S phase or mitotic phosphorylation sites act independently and have distinct functional roles.
Talasz
H
H
Institute of Medical Chemistry and Biochemistry, University of Innsbruck, Fritz-Pregl-Strasse 3, A-6020 Innsbruck, Austria.
Sapojnikova
N
N
Helliger
W
W
Lindner
H
H
Puschendorf
B
B
D016475:3T3 Cells; D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D002843:Chromatin / Q000502:physiology / Q000648:ultrastructure; D002851:Chromatography, High Pressure Liquid; D004279:DNA, Viral / Q000302:isolation & purification / Q000378:metabolism; D016193:G1 Phase; D006657:Histones / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008099:Liver / Q000648:ultrastructure / Q000821:virology; D008297:Male; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics*; D051379:Mice; D008938:Mitosis; D009707:Nucleosomes / Q000648:ultrastructure* / Q000821:virology*; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D020033:Protein Isoforms / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D016196:S Phase; D013737:Testis / Q000648:ultrastructure / Q000821:virology
D002843:Chromatin; D004279:DNA, Viral; D006657:Histones; D009707:Nucleosomes; D020033:Protein Isoforms