Characterization of a mutation in the acetolactate synthase of<i>Bacillus subtilis</i>that causes a cold-sensitive phenotype

Wiegeshoff, Frank; Marahiel, Mohamed A.

doi:10.1111/j.1574-6968.2007.00739.x

Cited by 9 publications

(10 citation statements)

References 27 publications

(37 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…subtilis strain AG174, our stock of strain JH642 (1). This widely used strain contains known point mutations conferring auxotrophy for tryptophan and phenylalanine and cold sensitivity (2, 3). We also report the sequence of AG1839 (also known as KPL69), a derivative of AG174 used in studies of replication initiation (4).…”

Section: Genome Announcementmentioning

confidence: 99%

Complete Genome Sequences of Bacillus subtilis subsp. subtilis Laboratory Strains JH642 (AG174) and AG1839

2014

View full text Add to dashboard Cite

show abstract

Section: Genome Announcementmentioning

confidence: 99%

Complete Genome Sequences of Bacillus subtilis subsp. subtilis Laboratory Strains JH642 (AG174) and AG1839

2014

View full text Add to dashboard Cite

show abstract

“…Osmoprotection and heat stress protection growth assays were conducted with B. subtilis wild-type laboratory strain JH642 and mutant derivatives thereof (Table 1). Cold stress protection growth assays, on the other hand, were conducted with B. subtilis laboratory strain 168 and mutant derivatives thereof (Table 1), since strain JH642 carries a mutation in the acetolactate synthase gene that confers a cold-sensitive growth phenotype (49). B. subtilis strains were routinely cultivated in Spizizen's minimal medium (SMM) with 0.5% (wt/vol) glucose as the carbon source, 15 mM (NH 4 ) 2 SO 4 as the nitrogen source, and L-tryptophan (20 mg liter Ϫ1 ) and L-phenylalanine (18 mg liter Ϫ1 ) to satisfy the auxotrophic requirements of B. subtilis strains JH642 (trpC2 pheA1) and 168 (trpC2), respectively (Table 1).…”

Section: Methodsmentioning

confidence: 99%

Dimethylglycine Provides Salt and Temperature Stress Protection to Bacillus subtilis

Bashir

Hoffmann

Smits

et al. 2014

Appl Environ Microbiol

View full text Add to dashboard Cite

e Glycine betaine is a potent osmotic and thermal stress protectant of many microorganisms. Its synthesis from glycine results in the formation of the intermediates monomethylglycine (sarcosine) and dimethylglycine (DMG), and these compounds are also produced when it is catabolized. Bacillus subtilis does not produce sarcosine or DMG, and it cannot metabolize these compounds. Here we have studied the potential of sarcosine and DMG to protect B. subtilis against osmotic, heat, and cold stress. Sarcosine, a compatible solute that possesses considerable protein-stabilizing properties, did not serve as a stress protectant of B. subtilis. DMG, on the other hand, proved to be only moderately effective as an osmotic stress protectant, but it exhibited good heat stress-relieving and excellent cold stress-relieving properties. DMG is imported into B. subtilis cells primarily under osmotic and temperature stress conditions via OpuA, a member of the ABC family of transporters. Ligand-binding studies with the extracellular solute receptor (OpuAC) of the OpuA system showed that OpuAC possesses a moderate affinity for DMG, with a K d value of approximate 172 M; its K d for glycine betaine is about 26 M. Docking studies using the crystal structures of the OpuAC protein with the sulfur analog of DMG, dimethylsulfonioacetate, as a template suggest a model of how the DMG molecule can be stably accommodated within the aromatic cage of the OpuAC ligand-binding pocket. Collectively, our data show that the ability to acquire DMG from exogenous sources under stressful environmental conditions helps the B. subtilis cell to cope with growth-restricting osmotic and temperature challenges.

show abstract

“…168 was generated by mutagenic X-rays and UV treatment of the wild type B. subtilis (Marburg) strain [2], resulting in the requirement for externally added tryptophan for growth, and the inability to produce a secreted antibiotic surfactin, due to mutations in the genes trpC and sfp , respectively [5]. Another broadly studied strain JH642 [6] which was obtained by multiple gene exchange experiments ([7], and James Hoch, personal communication) further differs from 168, including mutations in the genes pheA and ilvB that lead to phenylalanine requirement and cold sensitivity, respectively [8]. On the other hand, some laboratory strains (such as NCIB 3610 and SMY) do not have these phenotypes and are proposed to be true wild type strains.…”

Section: Introductionmentioning

confidence: 99%

High-Precision, Whole-Genome Sequencing of Laboratory Strains Facilitates Genetic Studies

et al. 2008

View full text Add to dashboard Cite

Whole-genome sequencing is a powerful technique for obtaining the reference sequence information of multiple organisms. Its use can be dramatically expanded to rapidly identify genomic variations, which can be linked with phenotypes to obtain biological insights. We explored these potential applications using the emerging next-generation sequencing platform Solexa Genome Analyzer, and the well-characterized model bacterium Bacillus subtilis. Combining sequencing with experimental verification, we first improved the accuracy of the published sequence of the B. subtilis reference strain 168, then obtained sequences of multiple related laboratory strains and different isolates of each strain. This provides a framework for comparing the divergence between different laboratory strains and between their individual isolates. We also demonstrated the power of Solexa sequencing by using its results to predict a defect in the citrate signal transduction pathway of a common laboratory strain, which we verified experimentally. Finally, we examined the molecular nature of spontaneously generated mutations that suppress the growth defect caused by deletion of the stringent response mediator relA. Using whole-genome sequencing, we rapidly mapped these suppressor mutations to two small homologs of relA. Interestingly, stable suppressor strains had mutations in both genes, with each mutation alone partially relieving the relA growth defect. This supports an intriguing three-locus interaction module that is not easily identifiable through traditional suppressor mapping. We conclude that whole-genome sequencing can drastically accelerate the identification of suppressor mutations and complex genetic interactions, and it can be applied as a standard tool to investigate the genetic traits of model organisms.

show abstract

Characterization of a mutation in the acetolactate synthase ofBacillus subtilisthat causes a cold-sensitive phenotype

Cited by 9 publications

References 27 publications

Complete Genome Sequences of Bacillus subtilis subsp. subtilis Laboratory Strains JH642 (AG174) and AG1839

Complete Genome Sequences of Bacillus subtilis subsp. subtilis Laboratory Strains JH642 (AG174) and AG1839

Dimethylglycine Provides Salt and Temperature Stress Protection to Bacillus subtilis

High-Precision, Whole-Genome Sequencing of Laboratory Strains Facilitates Genetic Studies

Contact Info

Product

Resources

About