Merck
  • Home
  • Search Results
  • Resuscitation-promoting factors are cell wall-lytic enzymes with important roles in the germination and growth of Streptomyces coelicolor.

Resuscitation-promoting factors are cell wall-lytic enzymes with important roles in the germination and growth of Streptomyces coelicolor.

Journal of bacteriology (2014-12-17)
Danielle L Sexton, Renée J St-Onge, Henry J Haiser, Mary R Yousef, Lauren Brady, Chan Gao, Jacqueline Leonard, Marie A Elliot
ABSTRACT

Dormancy is a common strategy adopted by bacterial cells as a means of surviving adverse environmental conditions. For Streptomyces bacteria, this involves developing chains of dormant exospores that extend away from the colony surface. Both spore formation and subsequent spore germination are tightly controlled processes, and while significant progress has been made in understanding the underlying regulatory and enzymatic bases for these, there are still significant gaps in our understanding. One class of proteins with a potential role in spore-associated processes are the so-called resuscitation-promoting factors, or Rpfs, which in other actinobacteria are needed to restore active growth to dormant cell populations. The model species Streptomyces coelicolor encodes five Rpf proteins (RpfA to RfpE), and here we show that these proteins have overlapping functions during growth. Collectively, the S. coelicolor Rpfs promote spore germination and are critical for growth under nutrient-limiting conditions. Previous studies have revealed structural similarities between the Rpf domain and lysozyme, and our in vitro biochemical assays revealed various levels of peptidoglycan cleavage capabilities for each of these five Streptomyces enzymes. Peptidoglycan remodeling by enzymes such as these must be stringently governed so as to retain the structural integrity of the cell wall. Our results suggest that one of the Rpfs, RpfB, is subject to a unique mode of enzymatic autoregulation, mediated by a domain of previously unknown function (DUF348) located within the N terminus of the protein; removal of this domain led to significantly enhanced peptidoglycan cleavage.

MATERIALS
Product Number
Brand
Product Description

Supelco
Chloramphenicol, VETRANAL®, analytical standard
Sigma-Aldrich
Chloramphenicol, ≥98% (HPLC)
Sigma-Aldrich
Chloramphenicol, meets USP testing specifications
Sigma-Aldrich
Ampicillin, anhydrous, 96.0-102.0% (anhydrous basis)
Sigma-Aldrich
Chloramphenicol, γ-irradiated
Sigma-Aldrich
Chloramphenicol, BioReagent, suitable for plant cell culture
Sigma-Aldrich
D-Cycloserine, synthetic
Sigma-Aldrich
Imidazole buffer Solution, BioUltra, 1 M in H2O
Sigma-Aldrich
Nitrilotriacetic acid disodium salt, Sigma Grade, ≥99%
Sigma-Aldrich
D-Cycloserine
Sigma-Aldrich
D-Cycloserine
Supelco
Ampicillin, analytical standard
Chloramphenicol, European Pharmacopoeia (EP) Reference Standard
USP
Ampicillin, United States Pharmacopeia (USP) Reference Standard
Ampicillin, anhydrous, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Sodium chloride, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
Sodium chloride solution, 0.9% in water, BioXtra, suitable for cell culture
Sigma-Aldrich
Sodium chloride, meets analytical specification of Ph. Eur., BP, USP, 99.0-100.5%
Sigma-Aldrich
Sodium chloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
Sodium chloride, tablet
Sigma-Aldrich
Sodium chloride, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
Imidazole, for molecular biology, ≥99% (titration)
Sigma-Aldrich
Imidazole, ≥99% (titration), crystalline
Ondansetron impurity E, European Pharmacopoeia (EP) Reference Standard
Supelco
Sodium chloride, Pharmaceutical Secondary Standard; Certified Reference Material
Imidazole, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Sodium chloride, Vetec, reagent grade, 99%
Sigma-Aldrich
Imidazole, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99%
SAFC
Sodium chloride solution, 5 M
Supelco
DL-Dithiothreitol solution, 1 M in H2O