Arnold J W, Shimkets L J
Department of Microbiology, University of Georgia, Athens 30602.
J Bacteriol. 1988 Dec;170(12):5765-70. doi: 10.1128/jb.170.12.5765-5770.1988.
The function of molecules associated with the cell surface may be determined by examining the phenotype of cells treated with inhibitors specific to these cell surface molecules. This strategy was used to examine the function of the major Congo red receptor of the myxobacterium Myxococcus xanthus, which has a developmental cycle that involves social interactions among cells. A class of social motility mutations (A+ S-), known as dsp, may inhibit the same subcellular component as Congo red because the phenotype of wild-type cells which had been treated with Congo red resembled in several ways the phenotype of the Dsp mutants. First, Congo red inhibited agglutination of wild-type cells, whereas Dsp cells were incapable of agglutinating, even in the absence of Congo red. Second, Congo red inhibited fruiting body formation by wild-type cells and reduced the yield of myxospores. Untreated Dsp cells were unable to form fruiting bodies and produced few myxospores. Third, Congo red reduced the rate of wild-type gliding motility to a level comparable to that of untreated Dsp cells, but did not inhibit the A motility of Dsp cells. Finally, binding studies showed that Dsp cells lacked the major Congo red receptor. Wild-type cells bound Congo red with an apparent association constant of 2.4 X 10(5) M-1, while Dsp cells bound it with an apparent association constant of 8.5 X 10(3) M-1. Binding of Congo red to wild-type cells was saturated in less than 10 min and was reversible when excess Congo red was removed. These results suggest that the Congo red receptors are controlled by the S motility system and that these receptors are involved in cell cohesion, social motility, and fruiting body formation.
与细胞表面相关的分子功能可以通过检测用针对这些细胞表面分子的抑制剂处理过的细胞的表型来确定。该策略被用于研究粘细菌黄色粘球菌主要刚果红受体的功能,黄色粘球菌具有涉及细胞间社会相互作用的发育周期。一类被称为dsp的社会运动性突变(A + S-)可能与刚果红抑制相同的亚细胞成分,因为用刚果红处理过的野生型细胞的表型在几个方面类似于Dsp突变体的表型。首先,刚果红抑制野生型细胞的凝集,而Dsp细胞即使在没有刚果红的情况下也无法凝集。其次,刚果红抑制野生型细胞的子实体形成并降低粘孢子的产量。未经处理的Dsp细胞无法形成子实体且产生的粘孢子很少。第三,刚果红将野生型滑动运动速率降低到与未经处理的Dsp细胞相当的水平,但不抑制Dsp细胞的A运动性。最后,结合研究表明Dsp细胞缺乏主要的刚果红受体。野生型细胞结合刚果红的表观缔合常数为2.4×10⁵ M⁻¹,而Dsp细胞结合它的表观缔合常数为8.5×10³ M⁻¹。刚果红与野生型细胞的结合在不到10分钟内达到饱和,并且当去除过量的刚果红时是可逆的。这些结果表明刚果红受体受S运动系统控制,并且这些受体参与细胞黏附、社会运动性和子实体形成。