Kyriukha Yevhenii, Watkins Maxwell B, Redington Jennifer M, Dastvan Reza, Uversky Vladimir N, Hopkins Jesse B, Pozzi Nicola, Korolev Sergey
Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, MO.
The Biophysics Collaborative Access Team (BioCat), Departments of Biology and Physics, Illinois Institute of Technology, Chicago, IL.
bioRxiv. 2024 May 29:2023.06.01.543259. doi: 10.1101/2023.06.01.543259.
The Partner and Localizer of BRCA2 (PALB2) is a scaffold protein that links BRCA1 with BRCA2 to initiate homologous recombination (HR). PALB2 interaction with DNA strongly enhances HR efficiency in cells. The PALB2 DNA-binding domain (PALB2-DBD) supports strand exchange, a complex multistep reaction conducted by only a few proteins such as RecA-like recombinases and Rad52. Using bioinformatics analysis, small-angle X-ray scattering, circular dichroism, and electron paramagnetic spectroscopy, we determined that PALB2-DBD is an intrinsically disordered region (IDR) forming compact molten globule-like dimer. IDRs contribute to oligomerization synergistically with the coiled-coil interaction. Using confocal single-molecule FRET we demonstrated that PALB2-DBD compacts single-stranded DNA even in the absence of DNA secondary structures. The compaction is bimodal, oligomerization-dependent, and is driven by IDRs, suggesting a novel strand exchange mechanism. Intrinsically disordered proteins (IDPs) are prevalent in the human proteome. Novel DNA binding properties of PALB2-DBD and the complexity of strand exchange mechanism significantly expands the functional repertoire of IDPs. Multivalent interactions and bioinformatics analysis suggest that PALB2 function is likely to depend on formation of protein-nucleic acids condensates. Similar intrinsically disordered DBDs may use chaperone-like mechanism to aid formation and resolution of DNA and RNA multichain intermediates during DNA replication, repair and recombination.
BRCA2的伴侣和定位蛋白(PALB2)是一种支架蛋白,它将BRCA1与BRCA2连接起来以启动同源重组(HR)。PALB2与DNA的相互作用可显著提高细胞中的HR效率。PALB2 DNA结合结构域(PALB2-DBD)支持链交换,这是一种仅由少数蛋白质(如RecA样重组酶和Rad52)进行的复杂多步反应。通过生物信息学分析、小角X射线散射、圆二色性和电子顺磁共振光谱,我们确定PALB2-DBD是一个形成紧密熔球样二聚体的内在无序区域(IDR)。IDR与卷曲螺旋相互作用协同促进寡聚化。使用共聚焦单分子荧光共振能量转移,我们证明即使在没有DNA二级结构的情况下,PALB2-DBD也能使单链DNA压缩。这种压缩是双峰的、依赖寡聚化的,并且由IDR驱动,提示了一种新的链交换机制。内在无序蛋白(IDP)在人类蛋白质组中普遍存在。PALB2-DBD的新型DNA结合特性和链交换机制的复杂性显著扩展了IDP的功能范围。多价相互作用和生物信息学分析表明,PALB2的功能可能依赖于蛋白质-核酸凝聚物的形成。类似的内在无序DBD可能利用类似伴侣的机制来辅助DNA复制、修复和重组过程中DNA和RNA多链中间体的形成和解析。