Metabolic Activities in Dormant Spores of Bacillus Species
Abstract
It has long been accepted that bacterial spores have no metabolic activity - this dogma has been brought into question by a published report providing evidence that there was massive rRNA and mRNA degradation and a significant level of transcription in dormant Bacillus subtilis spores. Research funded through a recent ARO STIR proposal to the Setlow group found evidence of slow metabolic activity. but only when the spores were held in certain media conditions. This proposal seeks to extend this initial study to establish the magnitude of this metabolism and its effect on dormant spore properties using spores of the species B. subtilis and B. megaterium. For experiments that measure catabolites during dormancy. Bacillus cell Jines that lack the germinant receptors will be used in order to prevent germination during the incubation of the spores. Identification and quantification and the fate of low molecular weight catabolites in spores while dormant and during germination will be analyzed using l 3C and 31 P-NMR and enzymatic assays for the presence of ATP; The fate of 3-phosphoglycerate to either acetate, pyruvate or glyceric acid will likewise be tracked by 31P and 13C-NMR; the fate of rRNA during dormant spore incubation will be assayed by extracting rRNA from purified spores and quantifying via separation and visualization on a polyacrylamide gel; transcriptomic analysis will be used to track mRNA transcription and degradation. For those conditions that allow for maximal dormant spore metabolic changes, the effects of such changes on spore properties = specifically resistance, germination and outgrowth = will be assayed. Resistance measurements include effects of wet heat, dry heat and UV radiation.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Jan 12, 2017
- Source ID
- W911NF1610024
Entities
People
- Peter Setlow
Organizations
- Army Contracting Command
- United States Army
- University of Connecticut Health Center