Perović-Ottstadt Sanja, Adell Teresa, Proksch Peter, Wiens Matthias, Korzhev Michael, Gamulin Vera, Müller Isabel M, Müller Werner E G
Institut für Physiologische Chemie, Abteilung Angewandte Molekularbiologie, Universität, Mainz, Germany.
Eur J Biochem. 2004 May;271(10):1924-37. doi: 10.1111/j.1432-1033.2004.04102.x.
Sponges (phylum Porifera) live in a symbiotic relationship with microorganisms, primarily bacteria. Until now, molecular proof for the capacity of sponges to recognize fungi in the surrounding aqueous milieu has not been available. Here we demonstrate, for the demosponge Suberites domuncula (Porifera, Demospongiae, Hadromerida), a cell surface receptor that recognizes (1-->3)-beta-D-glucans, e.g. curdlan or laminarin. This receptor, the (1-->3)-beta-D-glucan-binding protein, was identified and its cDNA analysed. The gene coding for the 45 kDa protein was found to be upregulated in tissue after incubation with carbohydrate. Simultaneously with the increased expression of this gene, two further genes showed an elevated steady state level of expression; one codes for a fibrinogen-like protein and the other for the epidermal growth factor precursor. Expression of the (1-->3)-beta-D-glucan-binding protein and the fibrinogen-like protein occurred in cells on the sponge surface, in the pinacoderm. By Western blotting, the product of the fibrinogen-like protein gene was identified, the recombinant protein isolated, and antibodies raised to this protein. Their application revealed that a 5 kDa factor is produced, which is apparently processed from the 77 kDa epidermal growth factor precursor. Finally, we provided evidence that a tyrosine kinase pathway is initiated in response to exposure to D-glucan; its phosphorylation activity could be blocked by aeroplysinin. In turn, the increased expression of the downstream genes was suppressed. We conclude that sponges possess a molecular mechanism for recognizing fungi via the d-glucan carbohydrates on their surfaces.
海绵动物(多孔动物门)与微生物(主要是细菌)存在共生关系。到目前为止,尚无分子证据表明海绵动物有能力识别周围水环境中的真菌。在此,我们证明了寻常海绵纲的皮海绵(多孔动物门、寻常海绵纲、硬海绵目)存在一种能识别(1→3)-β-D-葡聚糖(如可德胶或海带多糖)的细胞表面受体。该受体,即(1→3)-β-D-葡聚糖结合蛋白,已被鉴定并对其cDNA进行了分析。发现编码45 kDa蛋白的基因在与碳水化合物孵育后的组织中上调。在该基因表达增加的同时,另外两个基因的表达稳态水平也升高;一个编码类纤维蛋白原蛋白,另一个编码表皮生长因子前体。(1→3)-β-D-葡聚糖结合蛋白和类纤维蛋白原蛋白在海绵表面的细胞(扁平上皮)中表达。通过蛋白质免疫印迹法,鉴定了类纤维蛋白原蛋白基因的产物,分离了重组蛋白,并制备了针对该蛋白的抗体。它们的应用表明产生了一种5 kDa的因子,显然是从77 kDa的表皮生长因子前体加工而来。最后,我们提供证据表明,暴露于D-葡聚糖会引发酪氨酸激酶途径;其磷酸化活性可被刺参素阻断。反过来,下游基因的表达增加受到抑制。我们得出结论,海绵动物拥有一种通过其表面的d-葡聚糖碳水化合物识别真菌的分子机制。