Paidhungat M, Setlow P
Department of Biochemistry, University of Connecticut Health Center, Farmington, Connecticut, USA.
J Bacteriol. 2000 May;182(9):2513-9. doi: 10.1128/JB.182.9.2513-2519.2000.
Dormant Bacillus subtilis spores germinate in the presence of particular nutrients called germinants. The spores are thought to recognize germinants through receptor proteins encoded by the gerA family of operons, which includes gerA, gerB, and gerK. We sought to substantiate this putative function of the GerA family proteins by characterizing spore germination in a mutant strain that contained deletions at all known gerA-like loci. As expected, the mutant spores germinated very poorly in a variety of rich media. In contrast, they germinated like wild-type spores in a chemical germinant, a 1-1 chelate of Ca(2+) and dipicolinic acid (DPA). These observations showed that proteins encoded by gerA family members are required for nutrient-induced germination but not for chemical-triggered germination, supporting the hypothesis that the GerA family encodes receptors for nutrient germinants. Further characterization of Ca(2+)-DPA-induced germination showed that the effect of Ca(2+)-DPA on spore germination was saturated at 60 mM and had a K(m) of 30 mM. We also found that decoating spores abolished their ability to germinate in Ca(2+)-DPA but not in nutrient germinants, indicating that Ca(2+)-DPA and nutrient germinants probably act through parallel arms of the germination pathway.
休眠的枯草芽孢杆菌孢子在存在称为萌发剂的特定营养物质时会萌发。人们认为孢子通过由gerA操纵子家族编码的受体蛋白来识别萌发剂,该家族包括gerA、gerB和gerK。我们试图通过对一个在所有已知的类gerA位点都有缺失的突变菌株中的孢子萌发进行表征,来证实GerA家族蛋白的这种假定功能。正如预期的那样,突变孢子在各种丰富培养基中的萌发情况非常差。相比之下,它们在化学萌发剂(一种Ca²⁺与吡啶二羧酸(DPA)的1∶1螯合物)中能像野生型孢子一样萌发。这些观察结果表明,gerA家族成员编码的蛋白质是营养物诱导萌发所必需的,但不是化学触发萌发所必需的,这支持了GerA家族编码营养物萌发剂受体的假说。对Ca²⁺-DPA诱导萌发的进一步表征表明,Ca²⁺-DPA对孢子萌发的影响在60 mM时达到饱和,其米氏常数(Kₘ)为30 mM。我们还发现,去除孢子的包被会消除它们在Ca²⁺-DPA中萌发的能力,但不会影响在营养物萌发剂中的萌发能力,这表明Ca²⁺-DPA和营养物萌发剂可能通过萌发途径的平行分支起作用。