Wabiko H, Ochi K, Nguyen D M, Allen E R, Freese E
Laboratory of Molecular Biology, National Institute of Neurological and Communicative Disorders and Stroke, Bethesda, Maryland 20892.
J Bacteriol. 1988 Jun;170(6):2705-10. doi: 10.1128/jb.170.6.2705-2710.1988.
Three metE mutations of Bacillus subtilis, which cause cells to have a 25- to 200-fold decrease in L-methionine S-adenosyltransferase (EC 2.5.1.6) activity, were mapped between bioB and thr. The corresponding three metE mutants contained three- to fourfold less intracellular S-adenosylmethionine (SAM) but at least sevenfold more methionine than the metE+ strain when grown in synthetic medium. This indicates a strong feedback control of SAM on its synthesis. However, only the metE2 strain, with the lowest SAM concentration, grew at a slightly lower rate than the parent, which showed that an intracellular concentration of about 25 microM SAM was critical for growth at the normal rate. Neither DNA methylation (measured by bacteriophage luminal diameter 105 restriction) nor sporulation was affected at this low SAM concentration. Addition of methionine to the growth medium caused an increase in the pool of SAM in some but not all metE mutants. Coaddition of adenine did not change this result. However, the extent of sporulation (induced by mycophenolic acid) was decreased 50-fold in all mutants by the addition of methionine and adenine. Therefore, the combination of methionine and adenine suppresses sporulation regardless of whether it causes an increase in the level of SAM.
枯草芽孢杆菌的三个metE突变体,其导致细胞中L-甲硫氨酸S-腺苷甲硫氨酸转移酶(EC 2.5.1.6)活性降低25至200倍,被定位在bioB和thr之间。相应的三个metE突变体在合成培养基中生长时,细胞内S-腺苷甲硫氨酸(SAM)含量比metE+菌株少三到四倍,但甲硫氨酸含量至少多七倍。这表明SAM对其合成有很强的反馈控制。然而,只有SAM浓度最低的metE2菌株生长速度比亲本略低,这表明细胞内约25 microM的SAM浓度对于正常生长速度至关重要。在这种低SAM浓度下,DNA甲基化(通过噬菌体腔直径105限制测量)和芽孢形成均未受到影响。向生长培养基中添加甲硫氨酸会导致一些(但不是所有)metE突变体中SAM池增加。同时添加腺嘌呤不会改变这一结果。然而,添加甲硫氨酸和腺嘌呤后,所有突变体中(由霉酚酸诱导的)芽孢形成程度降低了50倍。因此,无论甲硫氨酸和腺嘌呤是否导致SAM水平升高,它们的组合都会抑制芽孢形成。