Stephens K, Hartzell P, Kaiser D
Department of Biochemistry, Stanford University, California 94305.
J Bacteriol. 1989 Feb;171(2):819-30. doi: 10.1128/jb.171.2.819-830.1989.
Mutants of Myxococcus xanthus that had lost the ability to glide were examined to elucidate the mechanism of gliding motility. Nonmotile mutants resulting from a single mutational step were all defective at the same locus, mgl, which implied an important role for the mgl product(s) in gliding. Deletion experiments, transposon insertion mutagenesis, and genetic rescue of mgl mutants mapped the locus to a 1.6-kilobase segment of Myxococcus DNA. Two species of RNA that hybridized with mgl DNA were found both during vegetative growth and during the starvation-induced development of fruiting bodies, which also requires cell movement. The two RNA species, of 1.5 and 1.3 kilobases, had the same 5' to 3' orientation and overlapped extensively. The DNA sequences of mgl+ and of seven mgl mutants were determined. Each mutant differed from mgl+ by a single-base-pair change in the sequence. Two adjacent open reading frames were found in the sequence hybridizing to both species of mgl RNA. Six of the single-base-pair changes, each of which would result in a single-amino-acid change, and an insertion-produced mgl mutation were located in the downstream open reading frame. This open reading frame (of 195 amino acids) is therefore an mgl gene, called mglA. The function of the upstream open reading frame is not known with certainty, although it does contain one of the mgl mutant sites and could be a second mgl gene.
为阐明滑行运动的机制,对丧失滑行能力的黄色粘球菌突变体进行了研究。由单个突变步骤产生的非运动突变体在同一基因座mgl均有缺陷,这表明mgl产物在滑行中起重要作用。缺失实验、转座子插入诱变以及mgl突变体的基因拯救将该基因座定位到黄色粘球菌DNA的一个1.6千碱基片段上。在营养生长期间以及饥饿诱导的子实体发育期间(这也需要细胞移动)均发现了两种与mgl DNA杂交的RNA。这两种RNA分别为1.5千碱基和1.3千碱基,具有相同的5'到3'方向且广泛重叠。测定了mgl +和七个mgl突变体的DNA序列。每个突变体与mgl +的序列差异均为单个碱基对的改变。在与两种mgl RNA杂交的序列中发现了两个相邻的开放阅读框。六个单个碱基对的改变(每个改变都会导致单个氨基酸的变化)以及一个插入产生的mgl突变位于下游开放阅读框中。因此,这个开放阅读框(由195个氨基酸组成)是一个mgl基因,称为mglA。上游开放阅读框的功能尚不确定,尽管它确实包含一个mgl突变位点,可能是第二个mgl基因。