Department of Microbiology and Immunobiology, Harvard Medical School , Boston, Massachusetts 02115, United States.
ACS Chem Biol. 2014 Feb 21;9(2):459-67. doi: 10.1021/cb4006744. Epub 2013 Nov 20.
Staphylococcus aureus is a Gram-positive pathogen with an unusual mode of cell division in that it divides in orthogonal rather than parallel planes. Through selection using moenomycin, an antibiotic proposed to target peptidoglycan glycosyltransferases (PGTs), we have generated resistant mutants containing a single point mutation in the active site of the PGT domain of an essential peptidoglycan (PG) biosynthetic enzyme, PBP2. Using cell free polymerization assays, we show that this mutation alters PGT activity so that much shorter PG chains are made. The same mutation in another S. aureus PGT, SgtB, has a similar effect on glycan chain length. Moenomycin-resistant S. aureus strains containing mutated PGTs that make only short glycan polymers display major cell division defects, implicating PG chain length in determining bacterial cell morphology and division site placement.
金黄色葡萄球菌是一种革兰氏阳性病原体,其细胞分裂方式不同寻常,它在垂直而不是平行的平面上分裂。通过使用莫能菌素(一种被认为靶向肽聚糖糖基转移酶(PGTs)的抗生素)进行选择,我们生成了含有单个点突变的耐药突变体,该突变体位于必需的肽聚糖(PG)生物合成酶 PBP2 的 PGT 结构域的活性位点中。使用无细胞聚合测定法,我们表明该突变改变了 PGT 活性,从而产生了更短的 PG 链。另一种金黄色葡萄球菌 PGT SgtB 中的相同突变对聚糖链长度有类似的影响。仅产生短聚糖聚合物的携带突变 PGT 的耐莫能菌素金黄色葡萄球菌菌株显示出主要的细胞分裂缺陷,这表明 PG 链长度决定了细菌细胞形态和分裂部位的放置。