Abstract：
FOXP2 is a transcription factor associated with speech and language. Like other FOX transcription factors, it has a DNA binding region called the forkhead domain (FHD). This domain can exist as a monomer or a domain swapped dimer. In addition to the FHD, the leucine zipper region (LZ) of FOXP2 is also believed to be associated with both DNA binding and oligomerization. To better understand the relationship between DNA binding and oligomerization of FOXP2, we investigated its structure, stability and dynamics, focusing specifically on the FHD and the LZ. We did this by using two constructs: one containing the isolated FHD and one containing both the LZ and the FHD (LZ-END). We demonstrate in this work, that while the FHD maintains a monomeric form that is capable of binding DNA, the LZ-END undergoes a dynamic transition between oligomeric states in the presence of DNA. Our findings suggest that FOXP2\'s LZ domain influences DNA binding affinity through a change in oligomeric state. We show through hydrogen exchange mass spectroscopy that certain parts of the FHD and interlinking region become less dynamic when in the presence of DNA, confirming DNA binding and oligomerization in these regions. Moreover, the detection of a stable equilibrium intermediate state during LZ-END unfolding supports the idea of cooperation between these two domains. Overall, our study sheds light on the interplay between two FOXP2 domains, providing insight into the protein\'s ability to respond dynamically to DNA, and enriching our understanding of FOXP2\'s role in gene regulation.
摘要：
FOXP2是与语音和语言相关的转录因子。像其他FOX转录因子一样，它有一个称为叉头域（FHD）的DNA结合区。该结构域可以作为单体或结构域交换二聚体存在。除了FHD,FOXP2的亮氨酸拉链区(LZ)也被认为与DNA结合和寡聚化相关。为了更好地理解FOXP2的DNA结合和寡聚化之间的关系，我们研究了其结构，稳定性和动力学，特别关注FHD和LZ。我们通过使用两个构建体:一个包含分离的FHD,一个包含LZ和FHD(LZ-END)。我们在这项工作中证明，虽然FHD保持能够结合DNA的单体形式，在DNA的存在下,LZ-END经历寡聚状态之间的动态转变。我们的发现表明FOXP2的LZ结构域通过寡聚状态的变化影响DNA结合亲和力。我们通过氢交换质谱显示，当存在DNA时，FHD和互连区域的某些部分变得不那么动态，确认这些区域中的DNA结合和寡聚化。此外，在LZ-END展开过程中检测到稳定的平衡中间状态支持这两个域之间合作的想法。总的来说,我们的研究揭示了两个FOXP2域之间的相互作用，深入了解蛋白质对DNA的动态反应能力，丰富了我们对FOXP2在基因调控中作用的认识。
