The binding of adenosine(5')tetraphospho(5')adenosine to calf thymus histones measured by non-equilibrium dialysis.
Binding of adenosine(5')tetraphospho(5')adenosine (Ap4A) to histones of calf thymus was investigated by non-equilibrium dialysis. Histone H1 interacts with the dinucleotide via two strong sites and competes with Mg2+ ions. Intrinsic dissociation constants were 1.6 +/- 0.1 microM and 11 +/- 1 microM for zero and 0.4 mm-Mg2+ concentration respectively. Binding of poly(dT) and of other nucleotides to histone H1 was measured in an [3H]Ap4A-competition assay. The tendency to form complexes among nucleotides was highest for bisnucleoside tetraphosphates and decreased in the order poly(dT) greater than or equal to Ap4A approximately Gp4G greater than Ap4 much greater than Ap3A approximately Ap5A greater than or equal to ATP, GTP and dTTP. The co-ordination complex derived from Ap4A and cis-diammine-dichloroplatinum(II) was not reactive. The other histones of calf thymus also bound Ap4A with affinities decreasing in the order H4 approximately H3 greater than H1 greater than H2b greater than H2a. Ap4A stimulated the exchange of histone H1 between nucleosomes, but this effect was referred to ionic strength. It did not bind to assembled nucleosomes. Binding of Ap4A to histone H1 was decreased by salt (NaCl). At physiological saline concentration the value of the dissociation constant is commensurable with the value of the Ap4A concentration in the nucleus and thus indicative of complex-formation in vivo.
Just
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Institut für Biophysik und Physikalische Biochemie der Universität, Regensburg, German Federal Republic.
Holler
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D000227:Adenine Nucleotides / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D002843:Chromatin / Q000378:metabolism; D003956:Dialysis; D015226:Dinucleoside Phosphates; D006657:Histones / Q000378:metabolism*; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D009711:Nucleotides / Q000378:metabolism; D009994:Osmolar Concentration; D011485:Protein Binding; D013950:Thymus Gland / Q000378:metabolism*
D000227:Adenine Nucleotides; D002843:Chromatin; D015226:Dinucleoside Phosphates; D006657:Histones; D046911:Macromolecular Substances; D009711:Nucleotides; C020733:diadenosine tetraphosphate; D008274:Magnesium