Sporulation in <i>Bacillus subtilis</i>. Correlation of biochemical events with morphological changes in asporogeneous mutants

Waites, W. M.; Kay, D.; Dawes, Ian W.; Wood, David А.; Warren, S. C.; Mandelstam, J.

doi:10.1042/bj1180667

Cited by 96 publications

(95 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The biochemical properties of the mutants with respect to production of enzymes associated with earlier stages of sporulation (Waites et al, 1970) was as expected; the three mutants all produced serine protease, alkaline phosphatase and glucose dehydrogenase. However, one of the three mutants, 516, was also defective in DPA production (DPA production is a stage V event).…”

Section: Discussionmentioning

confidence: 77%

Spovh And Spovj - New Sporulation Loci In BAcillus Subtilis 168

Hill

1983

Microbiology

View full text Add to dashboard Cite

Three sporulation mutants have been isolated which produce spores with an atypical resistance phenotype, i,e. they are sensitive to organic solvents and heat but resistant to lysozyme. All three mutants produced serine protease, alkaline phosphatase and glucose dehydrogenase which are biochemical marker events for stages I, I1 and 111. Two of the three mutants produced dipicolinic acid, a late marker, but the third was defective in its production. Heat-resistance was not restored to any of the mutants by the provision of exogenous dipicolinate. Gel electrophoresis showed that the mutant spores had similar patterns of spore coat proteins to the wild-type and electron microscopy revealed no significant structural differences. The three mutations responsible for the phenotypes of the mutant spores lie in the phe-argA region of the Bacillus subtilis chromosome. Recombination index values indicate that the mutations are in three separate genes. They define at least two new sporulation loci, designated spoVH and spo VJ. , 1980; Jenkinson et al., 1980, 1981). It should, therefore, be possible to isolate mutants in which a 'late' property is normal while an 'earlier' one is defective. We have now been able to show that mutants with this previously unexpected phenotype do exist and in this paper the isolation and characterization of three organic solvent-sensitive, heat-sensitive but lysozyme-resistant mutants are described. The mutations carried by these three mutants identify two new sporulation loci. METHODS Chemicals.Unless otherwise stated chemicals were obtained from Sigma. Organisms. Bacillus subtilis Marburg 168 trpC2 which requires either indole or tryptophan and which sporulates normally is referred to as the wild-type. Details of sporulation defective (Spo-) mutants derived from 168 and Spo+ auxotrophic strains used for genetic analysis are shown in Table 1

show abstract

Section: Discussionmentioning

confidence: 77%

Spovh And Spovj - New Sporulation Loci In BAcillus Subtilis 168

Hill

1983

Microbiology

View full text Add to dashboard Cite

show abstract

“…By electron microscopy, spo-54 was shown to be either a spu0 or a spu/ mutant since some cells were observed with slightly indented asymmetric septa. Various enzymes and compounds that are expressed at specific times during sporulation in B. subtilis (Waites et al, 1970) were tested in B. megateriurn QM B1551 to establish the kinetics of expression in this strain and to determine whether they might be used to help distinguish mutants blocked at specific stages during our characterization of the fusion mutants. In B. subtilis, alkaline phosphatase is expressed at t 2 , glucose dehydrogenase at t3 and DPA between t4 and t5 (Waites et al, 1970).…”

Section: Discussionmentioning

confidence: 99%

“…Synthesis of several enzymes such as alkaline phosphatase and glucose de hydrogenase begins during specific stages of sporulation in B. subtilis (Waites et al, 1970) and their synthesis has been used to help delineate mutants blocked at specific stages (Mandelstam & Errington, 1987;Sandman et al, 1987). Expression of these enzymes was tested in wild-type B. megaterium and compared to one of the mutants as shown in Fig.…”

Section: Enzyme Assaysmentioning

confidence: 99%

Isolation and characterization of sporulation lacZ fusion mutants of Bacillus megaterium

Tao

Vary

1991

Journal of General Microbiology

View full text Add to dashboard Cite

A derivative of Bacillus megaterium QM B1551 cured of all seven resident plasmids was mutated to Lac-. Transposon Tn917 carrying lac2 and cat was then used to isolate four spo: : fucZ-cat fusion mutants by screening for colonies expressing the fusion in stationary phase. The sporulation frequencies of the mutants ranged from lo-' to Macrolide, lincosamide and streptogramin B resistance (MLS') of the mutants cotransduced 100% with the sporulation defect and all four mutations mapped near trp by transduction in the order: trp-hisH-spo. All four mutants isolated were defective early in sporulation since P-galactosidase could be detected between zero and 2 h after the end of exponential growth. Electron microscopy of two of the mutants expressing the enzyme at t,-t2 revealed a defect prior to or just at the beginning of septum formation in one, and after completion of septum formation in the other. Little or no synthesis of dipicolinic acid, glucose dehydrogenase or alkaline phosphatase was detected in the mutants, but each was neutral protease positive. These results show that the mutations were in at least two early genes expressed before glucose dehydrogenase production. This study represents the first genetic characterization of sporulation mutants in B. megaterium and also demonstrates that gene fusion technology can be used in this species.

show abstract

“…phosphate repressible) enzyme (Warren 1968). Examination of the morphological and biochemical properties of asporogenous mutants suggested that this increase in phosphatase activity during spore formation was sporulationspecific and that it might be associated with stage II (spore septum formation) of the sporulation process (Waites et al 1970;Coote 1972). Cytochemical studies have provided additional evidence for linking the appearance of alkaline phosphatase during sporulation with stage II (Glenn 1971).…”

Section: Introductionmentioning

confidence: 99%

“…Early blocked sporulation mutants (i.e. all stage 0 and stage I and most of stage II) fail to produce alkaline phosphatase (Waites et al 1970;Coote 1972) although they produce normal levels of phosphatase when starved of phosphate in the vegetative state (Glenn 1971).…”

Section: Introductionmentioning

confidence: 99%

Alkaline Phosphatase Mutants of Bacillus subtilis

RGlenn¹

1975

Aust. Jnl. Of Bio. Sci.

View full text Add to dashboard Cite

Alkaline phosphatases from vegetative and sporulating cells of B. subtilis have been shown previously to be identical in all criteria examined. Despite this, 15 mutants producing low levels of the phosphatase during phosphate starvation of vegetative cells have been shown to produce high levels of the sporulation-specific alkaline phosphatase. It has been shown by imrnunochemical means that seven of these mutants when starved of phosphate produce low levels of normal wild-type enzyme. The sporulation form of the enzyme from one mutant (P-l00) has been shown to be identical with the phosphatases from vegetative and sporulating cells of the wild type. It is proposed that all the mutants have regulatory defects in the control of the alkaline phosphatase from vegetative cells but nevertheless retain an intact structural gene for the enzyme and the control system for the phosphatase during sporulation.

show abstract

Sporulation in Bacillus subtilis. Correlation of biochemical events with morphological changes in asporogeneous mutants

Cited by 96 publications

References 20 publications

Spovh And Spovj - New Sporulation Loci In BAcillus Subtilis 168

Spovh And Spovj - New Sporulation Loci In BAcillus Subtilis 168

Isolation and characterization of sporulation lacZ fusion mutants of Bacillus megaterium

Alkaline Phosphatase Mutants of Bacillus subtilis

Contact Info

Product

Resources

About