Archaebacterial histone-like protein MC1 can exhibit a sequence-specific binding to DNA.
The binding of MC1 protein, the major chromosomal protein of the archaebacterium Methanosarcina sp. CHTI 55, to the region preceding the strongly expressed genes encoding methyl coenzyme reductase in a closely related micro-organism has been investigated. By gel retardation and DNAase I footprinting assays, we identified a preferential binding sequence in an open reading frame of unknown function. The large area of DNA protected against DNAase I is interrupted by a strong cleavage enhancement site on each strand. By circular permutation assays, we showed that the DNA bends upon MC1 binding. Furthermore we observed that the presence of a sequence outside the binding site can induce an unusual electrophoretic behaviour in some complexes.
Centre de biophysique moléculaire, Orléans, France.
C
C
Teyssier
B
B
Laine
A
A
Gervais
J C
JC
Maurizot
F
F
Culard
D019843:Archaeal Proteins; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D004269:DNA, Bacterial / Q000737:chemistry / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D017020:Methanosarcina / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016366:Open Reading Frames / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics; D011485:Protein Binding; D015183:Restriction Mapping; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet
D019843:Archaeal Proteins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C051211:HMb protein, Methanosarcina; D012261:Ribonucleoproteins; D010088:Oxidoreductases; C021572:methyl coenzyme M reductase; D003850:Deoxyribonuclease I