Austin F E, Turco J, Winkler H H
Infect Immun. 1987 Jan;55(1):240-4. doi: 10.1128/iai.55.1.240-244.1987.
The growth requirement of Rickettsia prowazekii for the amino acids serine and glycine was assessed in both wild-type cell lines and a mutant cell line. X-irradiated L929 cells supported the growth of R. prowazekii when the cells were incubated in Eagle minimal essential medium supplemented with serum. In contrast, in this medium, X-irradiated Vero cells did not support the growth of rickettsiae unless cycloheximide, serine, or glycine was added. Other nonessential amino acids, additional glucose, and potential products of host cell metabolism of serine and glycine were nonstimulatory. The concentration of serine or glycine required to support rickettsial growth had no effect on the doubling time of uninfected, unirradiated Vero cells. A comparison of intracellular amino acid pools indicated that the serine and glycine concentrations in mock-infected Vero cells were approximately 31 and 14% of the respective concentrations in mock-infected L929 cells. The pools of both amino acids in Vero cells increased markedly upon treatment of the cells with cycloheximide. Interconversion of serine and glycine catalyzed by serine hydroxymethyltransferase was detected in cell-free extracts of purified rickettsiae. However, this enzymatic activity did not permit rickettsial growth in a glycine-requiring clone (772-56d) of the Chinese hamster ovary cell CHO-K1 in the absence of glycine supplementation. These data indicate that R. prowazekii depends on the host cell for serine or glycine.
在野生型细胞系和突变细胞系中评估了普氏立克次体对丝氨酸和甘氨酸的生长需求。当在补充了血清的伊格尔基本培养基中培养时,经X射线照射的L929细胞支持普氏立克次体的生长。相比之下,在这种培养基中,经X射线照射的Vero细胞除非添加放线菌酮、丝氨酸或甘氨酸,否则不支持立克次体的生长。其他非必需氨基酸、额外的葡萄糖以及丝氨酸和甘氨酸的宿主细胞代谢潜在产物均无刺激作用。支持立克次体生长所需的丝氨酸或甘氨酸浓度对未感染、未照射的Vero细胞的倍增时间没有影响。细胞内氨基酸库的比较表明, mock感染的Vero细胞中丝氨酸和甘氨酸的浓度分别约为mock感染的L929细胞中相应浓度的31%和14%。用放线菌酮处理细胞后,Vero细胞中这两种氨基酸的库均显著增加。在纯化的立克次体的无细胞提取物中检测到了由丝氨酸羟甲基转移酶催化的丝氨酸和甘氨酸的相互转化。然而,在没有补充甘氨酸的情况下,这种酶活性并不能使中国仓鼠卵巢细胞CHO-K1的甘氨酸需求克隆(772-56d)中的立克次体生长。这些数据表明,普氏立克次体的丝氨酸或甘氨酸依赖于宿主细胞。