MacNeil S D, Calara F, Hartzell P L
Department of Microbiology and Molecular Genetics, University of California, Los Angeles 90024-1489.
Mol Microbiol. 1994 Oct;14(1):61-71. doi: 10.1111/j.1365-2958.1994.tb01267.x.
Gliding is the directed movement of cells across surfaces which occurs in the absence of external organelles such as flagella. Gliding of the complex prokaryote, Myxococcus xanthus, results from the action of two independent sets of genes known as the A (adventurous motility) and S (social motility) genes. Strains with mutations in both systems (A-S-) do not spread on agar surfaces because both individual and group movement is abolished. To generate regulated, transcriptional fusions with operons including A and S genes, we introduced TN5-lac into A- and S- strains to obtain non-motile A-S::Tn5-lac and A::Tn5-lac S- double mutants. These insertions identify five separate clusters of A genes and three separate clusters of S genes on the M. xanthus genome. Some Tn5-lac insertions map near two of the five previously identified motility gene clusters, but at least five new clusters were identified in this search. Single mutations at only one locus, mglA, block motility; the mglA locus is epistatic to A and S motility genes. A- and S- Tn5-lac insertions were transduced into mgl+ and delta mgl strains. The levels of beta-galactosidase activity produced from each A- or S- Tn5-lac insertion are similar in otherwise isogenic mgl+ and delta mgl strains, showing that MglA does not regulate the transcription of many A and S genes.
滑行是细胞在没有鞭毛等外部细胞器的情况下在表面上的定向运动。复杂原核生物黄色黏球菌的滑行是由两组独立的基因,即A(冒险性运动)基因和S(社会性运动)基因的作用引起的。在这两个系统中都有突变的菌株(A-S-)不会在琼脂表面扩散,因为个体运动和群体运动都被消除了。为了产生与包括A和S基因的操纵子的受调控转录融合体,我们将TN5-lac引入A-和S-菌株中,以获得无运动能力的A-S::Tn5-lac和A::Tn5-lac S-双突变体。这些插入在黄色黏球菌基因组上鉴定出五个独立的A基因簇和三个独立的S基因簇。一些Tn5-lac插入位于五个先前鉴定的运动基因簇中的两个附近,但在这次搜索中至少鉴定出了五个新的簇。仅在一个位点mglA处的单突变就会阻断运动能力;mglA位点对A和S运动基因具有上位性。将A-和S- Tn5-lac插入物转导到mgl+和Δmgl菌株中。在其他方面同基因的mgl+和Δmgl菌株中,每个A-或S- Tn5-lac插入产生的β-半乳糖苷酶活性水平相似,这表明MglA不调节许多A和S基因的转录。