Graumann P, Marahiel M A
Philipps Universität Marburg, Germany.
FEBS Lett. 1994 Jan 31;338(2):157-60. doi: 10.1016/0014-5793(94)80355-2.
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.
我们已经对枯草芽孢杆菌主要冷休克蛋白CspB的核酸结合特性进行了表征。CspB是冷休克结构域（CSD）家族的成员，该家族在原核生物和真核生物中广泛存在，并共享核酸结合结构域CSD。CSD结构域高度保守，与Y-box基序具有很强的亲和力，Y-box基序是一种包含CTGATTGGC/TC/TAA序列的顺式元件。在一系列使用包含Y-box基序和改变序列的寡核苷酸的凝胶阻滞实验中，我们展示了CspB对包含ATTGG以及互补CCAAT Y-box核心序列的单链DNA的优先结合。相比之下，CspB对改变的Y-box核心序列表现出较低的亲和力。根据寡核苷酸的长度以及与Y-box核心序列的序列偏差程度，完全阻滞需要3至超过10倍过量的CspB。
