Identification of host genes that affect acquisition of an integrative and conjugative element in <scp><i>B</i></scp><i>acillus subtilis</i>

Johnson, Christopher M.; Grossman, Alan D.

doi:10.1111/mmi.12736

Cited by 57 publications

(71 citation statements)

References 89 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Previously, we found that mprF and several other genes involved in the synthesis of phospholipids affect the efficiency of conjugation (20). MprF catalyzes addition of a lysyl group from Lys-tRNA Lys to phosphatidylglycerol to form lysyl-phosphatidylglycerol ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The unmarked ⌬mprF459 allele was constructed by replacing mprF with cat flanked by lox sites to generate strain CMJ459 and then recombining out the lox-cat allele using Cre recombinase expressed from pDR244, as previously described (20,26). The genomic boundaries of this allele are the same as for the ⌬mprF125::mls and ⌬mprF162::spc alleles.…”

Section: Methodsmentioning

confidence: 99%

“…Mating assays were performed on filters as previously described (9,20). Briefly, donor and recipient cells were grown separately in minimal medium with 1% arabinose as a carbon source.…”

Section: Methodsmentioning

confidence: 99%

“…In Enterococcus faecalis, lipoteichoic acids may be important for mating pair formation (17)(18)(19). Recently, we found that in B. subtilis, the phospholipid head groups of the membrane bilayer make important contributions to conjugation (20).…”

mentioning

confidence: 99%

“…Previously, we used transposon insertions and deep sequencing (Tnseq) to identify genes in recipients that affect the frequency of conjugation (20). We found that deletion of genes involved in the synthesis of various phospholipids has distinct effects on the ability of B. subtilis to act as a recipient in conjugation.…”

mentioning

confidence: 99%

See 4 more Smart Citations

The Composition of the Cell Envelope Affects Conjugation in Bacillus subtilis

Johnson

Grossman

2016

J Bacteriol

Self Cite

View full text Add to dashboard Cite

Conjugation in bacteria is the contact-dependent transfer of DNA from one cell to another via donor-encoded conjugation machinery. It is a major type of horizontal gene transfer between bacteria. Conjugation of the integrative and conjugative element ICEBs1 into Bacillus subtilis is affected by the composition of phospholipids in the cell membranes of the donor and recipient. We found that reduction (or elimination) of lysyl-phosphatidylglycerol caused by loss of mprF caused a decrease in conjugation efficiency. Conversely, alterations that caused an increase in lysyl-phosphatidylglycerol, including loss of ugtP or overproduction of mprF, caused an increase in conjugation efficiency. In addition, we found that mutations that alter production of other phospholipids, e.g., loss of clsA and yfnI, also affected conjugation, apparently without substantively altering levels of lysyl-phosphatidylglycerol, indicating that there are multiple pathways by which changes to the cell envelope affect conjugation. We found that the contribution of mprF to conjugation was affected by the chemical environment. Wild-type cells were generally more responsive to addition of anions that enhanced conjugation, whereas mprF mutant cells were more sensitive to combinations of anions that inhibited conjugation at pH 7. Our results indicate that mprF and lysyl-phosphatidylglycerol allow cells to maintain relatively consistent conjugation efficiencies under a variety of ionic conditions. IMPORTANCEHorizontal gene transfer is a driving force in microbial evolution, enabling cells that receive DNA to acquire new genes and phenotypes. Conjugation, the contact-dependent transfer of DNA from a donor to a recipient by a donor-encoded secretion machine, is a prevalent type of horizontal gene transfer. Although critically important, it is not well understood how the recipient influences the success of conjugation. We found that the composition of phospholipids in the membranes of donors and recipients influences the success of transfer of the integrative and conjugative element ICEBs1 in Bacillus subtilis. Specifically, the presence of lysyl-phosphatidylglycerol enables relatively constant conjugation efficiencies in a range of diverse chemical environments.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Mating assays were performed on filters as previously described (9,20). Briefly, donor and recipient cells were grown separately in minimal medium with 1% arabinose as a carbon source.…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

The Composition of the Cell Envelope Affects Conjugation in Bacillus subtilis

Johnson

Grossman

2016

J Bacteriol

Self Cite

View full text Add to dashboard Cite

show abstract

Genetic Approaches to Uncover Gene Products Involved in Iron-Sulfur Protein Maturation: High-Throughput Genomic Screening Using Transposon Sequencing

Carabetta

Esquilin-Lebron

Zelzion

et al. 2021

Methods in Molecular Biology

View full text Add to dashboard Cite

Transposon sequencing: methods and expanding applications

Kwon

Ricke

Mandal

2015

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

A comprehensive understanding of genotype-phenotype links in bacteria is the primary theme of bacterial functional genomics. Transposon sequencing (Tn-seq) or its equivalent methods that combine random transposon mutagenesis and next-generation sequencing (NGS) represent a powerful approach to understand gene functions in bacteria on a genome-wide scale. This approach has been utilized in a variety of bacterial species to provide comprehensive information on gene functions related to various phenotypes or biological processes of significance. With further improvements in the molecular protocol for specific amplification of transposon junction sequences and increasing capacity of next generation sequencing technologies, the applications of Tn-seq have been expanding to tackle questions that are important yet difficult to address in the past. In this review, we will discuss the technical aspects of different Tn-seq methods along with their pros and cons to provide a helpful guidance for those who want to implement or improve Tn-seq for their own research projects. In addition, we also provide a comprehensive summary of recent published studies based on Tn-seq methods to give an updated perspective on the current and emerging applications of Tn-seq.

show abstract

Identification of host genes that affect acquisition of an integrative and conjugative element in Bacillus subtilis

Cited by 57 publications

References 89 publications

The Composition of the Cell Envelope Affects Conjugation in Bacillus subtilis

The Composition of the Cell Envelope Affects Conjugation in Bacillus subtilis

Genetic Approaches to Uncover Gene Products Involved in Iron-Sulfur Protein Maturation: High-Throughput Genomic Screening Using Transposon Sequencing

Transposon sequencing: methods and expanding applications

Contact Info

Product

Resources

About