Binding of Radioactive Benzylpenicillin to Sporulating
            <i>Bacillus</i>
            Cultures: Chemistry and Fluctuations in Specific Binding Capacity

Lawrence, Paul J.; Rogolsky, Marvin; Hanh, Vo Thi

doi:10.1128/jb.108.2.662-667.1971

Cited by 14 publications

(19 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In previous investigations (6,14), it was ob- added at stage 0 of spore formation. After 4 h, the bacteria were centrifuged and the cell-free supernatant was assayed for proteolytic and antibiotic activities as described in Materials and Methods.…”

Section: Resultsmentioning

confidence: 99%

“…Stage 2 of spore formation or septation is 57 on August 1, 2020 by guest http://jb.asm.org/ Downloaded from characterized by the formation of the inner forespore membrane which is believed to contain the germ cell wall synthesizing enzymes for peptidoglycan polymerization (14). The amount of penicillin that becomes bound to sporulating cells provides an estimate of the quantity of enzymes which function during the terminal steps of peptidoglycan synthesis (6,14). The penicillin-binding capacity of asporogenous mutants blocked during or prior to septation remains relatively constant or decreases during postlogarithmic growth (14).…”

mentioning

confidence: 99%

“…The penicillin-binding capacity of asporogenous mutants blocked during or prior to septation remains relatively constant or decreases during postlogarithmic growth (14). Alternatively, sporulating cells show two distinct intervals of enhanced binding capacity that occur during septation and cortex formation (6,14).…”

mentioning

confidence: 99%

“…In experiments designed to test the reversibility of EB inhibition, the agent was added to the cells at stage 0. After 3 h of incubation in EB the cells were washed with a tris(hydroxymethyl)aminomethanesuccinate-magnesium acetate buffer (TSM) (6), suspended in fresh SMM, and followed for the ability to produce refractile sporangia.…”

mentioning

confidence: 99%

“…The binding of ["4C]benzylpenicillin (Amersham-Searle) to treated and control organisms was determined by using a method previously described in this laboratory (6,14) with the exception that membrane filters containing cells with bound antibiotic were dissolved in a solution containing 1 ml of water and 9 ml of aquesol (New England Nuclear) prior to counting radioactivity. Cultures for the binding assays were inoculated, grown, and handled as described previously (6,14). The quantity of [14C]benzylpenicillin used to determine the specific binding capacity of sporulating cells was the minimal amount required for maximal binding under the given conditions.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Sensitivity of an Early Step in the Sporulation of Bacillus subtilis to Selective Inhibition by Ethidium Bromide

Rogolsky

Nakamura

1974

J Bacteriol

Self Cite

View full text Add to dashboard Cite

When a final concentration of 0.4 gg of ethidium bromide (EB) per ml, which is subinhibitory to vegetative growth, is added to sporulating cells of Bacillus subtilis Marburg during either stage 0 or the early part of stage 1, morphogenesis is blocked. If the given concentration of EB is added after the early part of stage 1, sporogenesis is unaffected. The synthesis of the serine protease and antibiotic, which are believed to be associated with sporulation events during the early part of stage 0, are not inhibited by EB. Enhanced binding of ["4C ]benzylpenicillin to sporulating cells during septation (stage 2) is a measure of the presence of terminal enzymes for germ cell wall peptidoglycan synthesis. EB does not interfere with the binding of penicillin to sporulating cells, but penicillin remains more permanently bound to EB-treated postlogarithmic cells than to untreated sporulating cells. The absence of an interval of increased penicillin binding activity during stage 2 by sporulating cells treated with EB indicates that EB blocks sporulation prior to the completion of the germ cell wall.The trypanocidal drug ethidium bromide (EB) (2, 7 diamino-9-phenyl-10-ethylphenanthidinium bromide) inhibits deoxyribonucleic acid (DNA) template function in a number of living organisms (11,12,19,20). The dye intercalates between DNA base pairs and prevents the initiation of ribonucleic acid (RNA) chains catalyzed by DNA-dependent RNA polymerase (11,12,20). If certain plasmid-positive bacterial cells are grown in the presence of EB, convalently-closed circular extrachromosomal DNA can become defective in the ability to replicate and be eliminated from growing cells (1,2,13,15). In general, covalently-closed circular templates are more sensitive to inactivation by EB than linear templates, although considerable variation in sensitivity to the dye exists with both types of templates (12). The variation in template sensitivities is believed to be dependent on the binding affinity of EB to particular initiation sites used by RNA polymerase (11,12,16,20).Sankaran and Pogell (16) recently reported that the promoter regions for catabolite-repressible genes on the Escherichia coli chromosome are particularly sensitive to inhibition by EB. Bacterial sporulation is a catabolite-repressible process (3,17), and the purpose of this investigation was to examine whether the stages of sporogenesis in Bacillus subtilis show selective inhibition to EB.The sporulation process involves an orderly sequence of seven clearly-defined morphological stages (14). The structural modification of vegetative RNA polymerase is believed to represent a vital control mechanism for the initiation of sporogenesis (3). Early genes for sporulation can then be recognized by the altered RNA polymerase. Greenleaf and Losick (4) suggested that the sporulation RNA polymerase of B. subtilis is modified by the loss of vegetative sigma factor and by the binding to a 70,000-dalton protein. Alternatively, Leighton et al. (7,8) believe that a postlogarithmic serine protease ...

show abstract

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations