SEMBID,s.r.o.-Research Centre of Applied Biomedical Diagnostics, Magnezitarska 2/C, 04013, Kosice, Slovakia.
Department of Cellular Logistics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077, Goettingen, Germany.
J Mol Evol. 2019 Sep;87(7-8):221-230. doi: 10.1007/s00239-019-09904-5. Epub 2019 Aug 12.
Our previous calculations of ionic interactions indicated that the Caenorhabditis elegans intermediate filament (IF) IFA proteins, in addition to IFA/IFB-1 heterodimers, may also form homodimers. In order to prove the significance of these calculations, we analysed the dimerization potential of the IFA chains in blot overlays. Unexpectedly, we found here that the dimerization of the IFA-1 protein was of both homotypic and heterotypic nature, and involved all proteins immobilized on the membrane (IFA-1, IFA-2, IFA-4, IFB-1, IFB-2, IFC-1, IFC-2, IFD-1, IFD-2 and IFP-1). A similar interaction profile, though less complex, was observed for two biotinylated proteins (IFA-2 and IFA-4). These and previous results indicate that the IFA proteins are able to form many different heteropolymeric and homopolymeric complexes in the C. elegans tissue, but that only those triggered by the IFA-specific IFB-1 protein result in mature IFs. Moreover, the calculations of the possible ionic interactions between the individual rod sequences as well as their various deletion variants indicated a special role in this process for the middle part of the C. elegans IF coil 1B segment that is deleted in all vertebrate cytoplasmic IFs. We hypothesized here, therefore, that the striking promiscuity of the C. elegans IFs originally involved a nuclear lamin which, due to a two-heptad-long rod deletion, prevented formation of a functional lamin/cIF dimer. This, in concert with an efficient dimerization and a strict tissue-specific co-expression, may allow expansion and maintenance of the multiple Caenorhabditis IFs. A possible implication for evolution of chordate IFs proteins is also discussed.
我们之前的离子相互作用计算表明,除了 IFA/IFB-1 异二聚体外,秀丽隐杆线虫中间丝 (IF) 的 IFA 蛋白也可能形成同二聚体。为了证明这些计算的重要性,我们在印迹杂交中分析了 IFA 链的二聚化潜力。出乎意料的是,我们在这里发现 IFA-1 蛋白的二聚化具有同型和异型两种性质,涉及固定在膜上的所有蛋白质 (IFA-1、IFA-2、IFA-4、IFB-1、IFB-2、IFC-1、IFC-2、IFD-1、IFD-2 和 IFP-1)。两种生物素化蛋白 (IFA-2 和 IFA-4) 的相互作用模式虽然不那么复杂,但也观察到了类似的情况。这些和之前的结果表明,IFA 蛋白能够在秀丽隐杆线虫组织中形成许多不同的异多聚体和同多聚体复合物,但只有那些由 IFA 特异性的 IFB-1 蛋白触发的复合物才能形成成熟的 IF。此外,对个别杆序列及其各种缺失变体之间可能的离子相互作用的计算表明,在这个过程中,在所有脊椎动物细胞质 IF 中缺失的秀丽隐杆线虫 IF 螺旋 1B 段的中间部分起着特殊的作用。因此,我们在这里假设,秀丽隐杆线虫 IF 的惊人混杂性最初涉及核层粘连蛋白,由于两个七肽长的杆缺失,阻止了功能性核层粘连蛋白/cIF 二聚体的形成。这与有效的二聚化和严格的组织特异性共表达相结合,可能允许扩展和维持多个秀丽隐杆线虫 IF。还讨论了对脊索动物 IF 蛋白进化的可能影响。