Differentiation by magnesium ions of affinities of nuclear proteins for consensus core nucleotide element of the transcription factor c-Myc in murine brain.
The addition of divalent cations such as Mg(2+) and Ca(2+) ions markedly reduced binding of a radiolabeled double stranded oligonucleotide probe for the transcription factor c-Myc in the presence of 100 mM KCl in nuclear extracts of the mouse whole brain. Irrespective of the addition of MgCl(2), binding was selectively competed with the unlabeled probe for c-Myc having a double stranded conformation. Treatment with V8 protease differentially modulated binding of the probe for c-Myc determined in the presence and absence of added MgCl(2). Introduction of irreversible covalent bonding between the radiolabeled probe and nuclear proteins led to retarded mobility of the radioactive probe/protein complex in the presence of MgCl(2) on sodium dodecyl sulfate electrophoresis regardless of treatment with DNase. However, an antibody against the c-Myc protein affected neither mobility nor intensity of the radioactive band on gel retardation electrophoresis. Moreover, regional distribution was different from each other in mouse brain when determined in the presence and absence of added MgCl(2). These results suggest that magnesium ions may have an ability to differentiate between nuclear c-Myc family proteins with different affinities for the consensus core nucleotide element CACGTG in murine brain.
Ogita
K
K
Department of Pharmacology, Setsunan University, Hirakata, Osaka, Japan.
Amizuka
T
T
Azuma
Y
Y
Yoneda
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism*; D016384:Consensus Sequence; D011756:Diphosphates / Q000378:metabolism; D006868:Hydrolysis; D008274:Magnesium / Q000378:metabolism*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009687:Nuclear Proteins / Q000378:metabolism*; D015345:Oligonucleotide Probes; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D014466:Ultraviolet Rays
D000911:Antibodies, Monoclonal; D011756:Diphosphates; D009687:Nuclear Proteins; D015345:Oligonucleotide Probes; D016271:Proto-Oncogene Proteins c-myc; D014157:Transcription Factors; D008274:Magnesium; C003319:sodium pyrophosphate