EXPLORING THE BINDING AFFINITIES OF C-JUN HOMODIMER TO THE SINGLE-NUCLEOTIDE MUTANT AP1 CONSENSUS SITES WITH DNA MICROARRAY
AP1 famity proteins regulate the expression of a myriad of genes in a variety of tissues and cell types. But in naturalenhancer regions, the sequences of AP1 regulatory elements often deviate from the optimal recognition consensus sequence TGA (C/G) TCA, To investigate the binding interaction of AP1 with those regulatory elements deviated from the optimal recognition consensus, we fabricated a double-stranded DNA (dsDNA) microarray with all possible single-nucleotide mutated (SNM) sequences of AP1 consensus site and explored the binding affinity of these SNM sequences with one of AP1 family protein, c-Jun. The importance of each nucleotide of AP1 site for the sequence-specific DNA-protein interaction was thus evaluated. The T5, C6, and A7 are most important for Jun-JunAP1 binding interaction and determine the specificity of interaction, the replacements to any other nucleotide could result in the similarly binding affinity losses. Comparatively, the T1, G2, A3, and G4 are less important for the binding affinity, the substitutions of these nucleotides could change the binding affinity differently. The G4 is least important for interaction, the randomized nucleotide exchange to G4 results increasing binding affinity. Among all possible single-nucleotide mutants, the A3 to C mutation has the highest binding affinity. These results can be cross-validated by the findings from crystallography and EMSA studies on DNA/protein interaction. Our studies demonstrate that the nucleotides at different positions contribute differently to the binding interaction.
G-Jun homodimer binding affinity single-nucleotide mutant AP1 consensus sites
Minli Li Donghua Zuhong Lu Jinke Wang
State Key Laboratory of Bioelectronics, Southeast University,Nanjing 210096, China
国际会议
The 4th International Forum on Post-genome Technologies(4IFPT)(第四届国际后基因组生命科学技术学术论坛)
杭州
英文
319-321
2006-09-25（万方平台首次上网日期，不代表论文的发表时间）