The major cold shock protein of Bacillus subtilis CspB binds with high affinity to the ATTGG- and CCAAT sequences in single stranded oligonucleotides.
We have characterized the nucleic acid binding properties of the major cold shock protein of Bacillus subtilis, CspB. CspB is a member of the cold shock domain (CSD) family, which is widespread among pro- and eukaryotes and shares the nucleic acid binding domain CSD. The CSD domain is highly conserved and binds with strong affinity to the Y-box motif, a cis-element that contains the CTGATTGGC/TC/TAA sequence. In a series of gel retardation experiments using oligonucleotides, which contain the Y-box motif and altered sequences, we show the preferential binding of CspB to single-stranded DNA that contains the ATTGG as well as the complementary CCAAT Y-box core sequence. In contrast CspB exhibits lower affinity to altered Y-box core sequences. Dependent on the length of the oligonucleotide and the degree of sequence deviation from the Y-box core sequence 3- to over 10-fold overexcess of CspB was needed for complete retardation.
Graumann
P
P
Philipps Universität Marburg, Germany.
Marahiel
M A
MA
D001412:Bacillus subtilis / Q000737:chemistry*; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism*
D001426:Bacterial Proteins; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D009841:Oligonucleotides; C077188:cold-shock protein CspB, Bacteria