A group of chromosomal proteins is specifically released by spermine and loses DNA-binding activity upon phosphorylation.
Biologically relevant concentrations as low as 500 microM spermine led to the specific release of chromatin-associated proteins from nuclei of rice (Oryza sativa) seedlings. Using a southwestern technique, it was shown that several of these proteins bind DNA. This affinity was lost upon in organello phosphorylation by an endogenous kinase. The effect of spermine was very specific. Spermidine was far less effective and putrescine was essentially ineffective in releasing these proteins. The most abundant spermine-released protein was shown to be homologous to the maize HMG1 protein. Our results suggest that spermine induces the release of spermine-released proteins by changing DNA conformation. Binding of these proteins might be sensitive to long-range changes in chromosome structure caused by torsional stress.
Van den Broeck
D
D
Laboratorium voor Genetica, Universiteit Gent, Belgium.
Van der Straeten
D
D
Van Montagu
M
M
Caplan
A
A
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002843:Chromatin / Q000187:drug effects / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases; D006609:High Mobility Group Proteins / Q000302:isolation & purification / Q000378:metabolism; D006657:Histones / Q000302:isolation & purification / Q000378:metabolism; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D012275:Oryza / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010766:Phosphorylation; D011700:Putrescine / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D013095:Spermidine / Q000494:pharmacology; D013096:Spermine / Q000494:pharmacology*
D002843:Chromatin; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D006657:Histones; D009687:Nuclear Proteins; D010446:Peptide Fragments; D013096:Spermine; D000255:Adenosine Triphosphate; D010450:Endopeptidases; D013095:Spermidine; D011700:Putrescine