DNA replication and cell enlargement of Enterococcus faecalis protoplasts

Kami, Satoshi; Tsuchikado, Rintaro; Nishida, Hiromi

doi:10.3934/microbiol.2019.4.347

Cited by 7 publications

(24 citation statements)

References 32 publications

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“…Both cell enlargement and DNA replication stop at 120 h of incubation, with no further change in cell size and DNA amount [ 74 ]. In order to investigate whether DNA replication influences bacterial protoplast or spheroplast enlargement, the DNA replication inhibitor novobiocin [ 75 , 76 ] was added to the incubation medium during the enlargement of E. faecalis protoplasts [ 74 , 77 ]. As a result, novobiocin inhibited not only DNA replication but also cell enlargement [ 74 , 77 ].…”

Section: Chromosomal Dna (Replication)mentioning

confidence: 99%

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Factors That Affect the Enlargement of Bacterial Protoplasts and Spheroplasts

Nishida

2020

IJMS

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Cell enlargement is essential for the microinjection of various substances into bacterial cells. The cell wall (peptidoglycan) inhibits cell enlargement. Thus, bacterial protoplasts/spheroplasts are used for enlargement because they lack cell wall. Though bacterial species that are capable of gene manipulation are limited, procedure for bacterial cell enlargement does not involve any gene manipulation technique. In order to prevent cell wall resynthesis during enlargement of protoplasts/spheroplasts, incubation media are supplemented with inhibitors of peptidoglycan biosynthesis such as penicillin. Moreover, metal ion composition in the incubation medium affects the properties of the plasma membrane. Therefore, in order to generate enlarged cells that are suitable for microinjection, metal ion composition in the medium should be considered. Experiment of bacterial protoplast or spheroplast enlargement is useful for studies on bacterial plasma membrane biosynthesis. In this paper, we have summarized the factors that influence bacterial cell enlargement.

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Section: Chromosomal Dna (Replication)mentioning

confidence: 99%

“…Although mitomycin C degrades chromosomal DNA of E. faecalis protoplasts, novobiocin does not [ 74 ]. When novobiocin was removed from the medium, E. faecalis protoplasts enlarged again [ 74 , 77 ]. Thus, DNA replication is associated with the biosynthesis of the plasma membrane.…”

Section: Chromosomal Dna (Replication)mentioning

confidence: 99%

Factors That Affect the Enlargement of Bacterial Protoplasts and Spheroplasts

Nishida

2020

IJMS

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“…It has been reported that the bacterial chromosome DNA attaches to the plasma membrane [ 5 – 10 ] although cell division does not occur in bacterial protoplasts or spheroplasts in the presence of an inhibitor of peptidoglycan biosynthesis [ 11 – 14 ]; removal of the peptidoglycan synthetic inhibitor can induce recovery from Escherichia coli spheroplasts to their native forms [ 15 – 17 ]. However, spheroplasts can enlarge in Difco Marine Broth (DMB) containing a peptidoglycan biosynthesis inhibitor, for example, penicillin [ 18 – 21 ], and interestingly, DNA replication has been reported during protoplast or spheroplast enlargement [ 13 , 14 , 18 , 19 , 23 ]. In the protoplasts of the gram-positive lactic acid bacterium Enterococcus faecalis , the amount of DNA increased during the protoplast enlargement between 0 and 96 h of incubation [ 23 ].…”

Section: Introductionmentioning

confidence: 99%

“…However, spheroplasts can enlarge in Difco Marine Broth (DMB) containing a peptidoglycan biosynthesis inhibitor, for example, penicillin [ 18 – 21 ], and interestingly, DNA replication has been reported during protoplast or spheroplast enlargement [ 13 , 14 , 18 , 19 , 23 ]. In the protoplasts of the gram-positive lactic acid bacterium Enterococcus faecalis , the amount of DNA increased during the protoplast enlargement between 0 and 96 h of incubation [ 23 ]. In addition, when the DNA replication inhibitor novobiocin was added to the medium after 24 h of incubation, the E. faecalis protoplast enlargement, as well as DNA replication was inhibited.…”

Section: Introductionmentioning

confidence: 99%

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Novobiocin inhibits membrane synthesis and vacuole formation of Enterococcus faecalis protoplasts

et al. 2020

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We demonstrate that plasma membrane biosynthesis and vacuole formation require DNA replication in Enterococcus faecalis protoplasts. The replication inhibitor novobiocin inhibited not only DNA replication but also cell enlargement (plasma membrane biosynthesis) and vacuole formation during the enlargement of the E. faecalis protoplasts. After novobiocin treatment prior to vacuole formation, the cell size of E. faecalis protoplasts was limited to 6 μm in diameter and the cells lacked vacuoles. When novobiocin was added after vacuole formation, E. faecalis protoplasts grew with vacuole enlargement; after novobiocin removal, protoplasts were enlarged again. Although cell size distribution of the protoplasts was similar following the 24 h and 48 h novobiocin treatments, after 72 h of novobiocin treatment there was a greater number of smaller sized protoplasts, suggesting that extended novobiocin treatment may inhibit the re-enlargement of E. faecalis protoplasts after novobiocin removal. Our findings demonstrate that novobiocin can control the enlargement of E. faecalis protoplasts due to inhibition of DNA replication.

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Effects of heterologous genome microinjection on the enlargement of Enterococcus faecalis protoplasts

Takahashi

Nishida

2022

Current Research in Microbial Sciences

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Highlights More than 100 E. faecalis protoplasts were generated and microinjected into heterologous genomic DNAs. Protoplast enlargement did not stop but continued after the microinjection. Microinjection of genomic DNA into E. faecalis protoplasts frequently induced weakness in enlargement.

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DNA replication and cell enlargement of Enterococcus faecalis protoplasts

Cited by 7 publications

References 32 publications

Factors That Affect the Enlargement of Bacterial Protoplasts and Spheroplasts

Factors That Affect the Enlargement of Bacterial Protoplasts and Spheroplasts

Novobiocin inhibits membrane synthesis and vacuole formation of Enterococcus faecalis protoplasts

Effects of heterologous genome microinjection on the enlargement of Enterococcus faecalis protoplasts

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