Molecular cloning, sequencing, and expression of lytM, a unique autolytic gene of Staphylococcus aureus

Ramadurai, Lakshmi; Jayaswal, Radheshyam K.

doi:10.1128/jb.179.11.3625-3631.1997

Cited by 79 publications

(91 citation statements)

References 33 publications

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“…Of the 12 genes potentially regulated by the YycG/YycF system listed in Table 2, 5 have been described as pathogenicity factors: 2 encode major antigens, SsaA and IsaA, and 3 others encode bacterial proteins implicated in interactions with the host matrix (a sialoproteinbinding protein, an elastin-binding protein, and a staphylokinase precursor). One gene, lytM, encodes a peptidoglycan hydrolase, a function shown to be regulated by the YycG/YycF system in B. subtilis (18,33). We note that 3 of the 19 genes of unknown function preceded by a potential YycF binding site encode proteins with significant similarities to the SsaA antigen (SA0620, SA2097, and SA2353).…”

Section: Resultsmentioning

confidence: 99%

“…In S. aureus, the present work has highlighted binding of YycF to the lytM promoter region. LytM is a glycylglycine endopeptidase (33,34), indicating a likely role for the S. aureus YycG/YycF system in controlling cell wall metabolism, as shown for its ortholog in B. subtilis. The yycF operon is transcribed during exponential growth in B. subtilis, and its expression is rapidly shut down as cells enter stationary phase (11).…”

Section: See Materials and Methods) His-tagged Yycf Andmentioning

confidence: 99%

“…In addition to ssaA, four genes closely linked to S. aureus virulence have the YycF consensus sequence in their promoter regions: sdrD, encoding a sialoprotein-binding protein; ebpS, encoding an elastin-binding protein; sak, encoding a staphylokinase precursor; and isaA, encoding immunodominant antigen A (Table 2). Furthermore, as described in B. subtilis (18), it seems that the YycG/YycF system also regulates cell wall metabolism in S. aureus via regulation of lytM, which encodes a peptidoglycan hydrolase (33). DNase I footprinting assays were performed on radiolabeled DNA fragments corresponding to the lytM and isaA promoter regions, generated by PCR with oligonucleotides OSA92/OSA93 (isaA) and OSA90/OSA91 (lytM) (the oligonucleotide sequences are listed in Table 1).…”

Section: See Materials and Methods) His-tagged Yycf Andmentioning

confidence: 99%

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Identification of Genes Controlled by the Essential YycG/YycF Two-Component System ofStaphylococcus aureus

2004

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Section: Resultsmentioning

confidence: 99%

Section: See Materials and Methods) His-tagged Yycf Andmentioning

confidence: 99%

Section: See Materials and Methods) His-tagged Yycf Andmentioning

confidence: 99%

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Identification of Genes Controlled by the Essential YycG/YycF Two-Component System ofStaphylococcus aureus

2004

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“…The C-terminal domains of the four proteins belong to COG4942 (membrane-bound metallopeptidases, involved in cell division and chromosome partitioning) and are characteristic for members of the M23/M37 family (Pfam01551) of putative zinc metalloendopeptidases from Gram-negative and Gram-positive bacteria (http://www.sanger.ac.uk/Software/Pfam/). The M37 protease family includes staphylococcal glycylglycine endopeptidases that hydrolyse the polyglycine interpeptide bridges of peptidoglycan of Gram-positive bacteria (Ramadurai & Jayaswal, 1997;Ramadurai et al, 1999;Recsei et al, 1987;Sugai et al, 1997) and the E. coli lipoprotein Fig. 7.…”

Section: Discussionmentioning

confidence: 99%

The amoebae plate test implicates a paralogue of lpxB in the interaction of Legionella pneumophila with Acanthamoeba castellanii

et al. 2005

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Legionella pneumophila is a bacterial parasite of freshwater amoebae which also grows in alveolar macrophages and thus causes the potentially fatal pneumonia Legionnaires' disease. Intracellular growth within amoebae and macrophages is mechanistically similar and requires the Icm/Dot type IV secretion system. This paper reports the development of an assay, the amoebae plate test (APT), to analyse growth of L. pneumophila wild-type and icm/dot mutant strains spotted on agar plates in the presence of Acanthamoeba castellanii. In the APT, wild-type L. pneumophila formed robust colonies even at high dilutions, icmT, -R, -P or dotB mutants failed to grow, and icmS or -G mutants were partially growth defective. The icmS or icmG mutant strains were used to screen an L. pneumophila chromosomal library for genes that suppress the growth defect in the presence of the amoebae. An icmS suppressor plasmid was isolated that harboured the icmS and flanking icm genes, indicating that this plasmid complements the intracellular growth defect of the mutant. In contrast, different icmG suppressor plasmids rendered the icmG mutant more cytotoxic for A. castellanii without enhancing intracellular multiplication in amoebae or RAW264.7 macrophages. Deletion of individual genes in the suppressor plasmids inserts identified lcs (Legionella cytotoxic suppressor) -A, -B, -C and -D as being required for enhanced cytotoxicity of an icmG mutant strain. The corresponding proteins show sequence similarity to hydrolases, NlpD-related metalloproteases, lipid A disaccharide synthases and ABC transporters, respectively. Overexpression of LcsC, a putative paralogue of the lipid A disaccharide synthase LpxB, increased cytotoxicity of an icmG mutant but not that of other icm/dot or rpoS mutant strains against A. castellanii. Based on sequence comparison and chromosomal location, lcsB and lcsC probably encode enzymes involved in cell wall maintenance and peptidoglycan metabolism. The APT established here may prove useful to identify other bacterial factors relevant for interactions with amoeba hosts.

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“…32 RNAIII, on the other hand, is predominantly expressed at high cell density, i.e. during the stationary phase of growth 27 and was proposed to control the synthesis of LytM at the translational level.…”

Section: Two Lytm Transcripts Originate From Different Promotersmentioning

confidence: 99%

Various checkpoints prevent the synthesis ofStaphylococcus aureuspeptidoglycan hydrolase LytM in the stationary growth phase

et al. 2016

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In Staphylococcus aureus, peptidoglycan metabolism plays a role in the host inflammatory response and pathogenesis. Transcription of the peptidoglycan hydrolases is activated by the essential 2-component system WalKR at low cell density. During stationary growth phase, WalKR is not active and transcription of the peptidoglycan hydrolase genes is repressed. In this work, we studied regulation of expression of the glycylglycine endopeptidase LytM. We show that, in addition to the transcriptional regulation mediated by WalKR, the synthesis of LytM is negatively controlled by a unique mechanism at the stationary growth phase. We have identified 2 different mRNAs encoding lytM, which vary in the length of their 5 0 untranslated (5 0 UTR) regions. LytM is predominantly produced from the WalKR-regulated mRNA transcript carrying a short 5 0 UTR. The lytM mRNA is also transcribed as part of a polycistronic operon with the upstream SA0264 gene and is constitutively expressed. Although SA0264 protein can be synthesized from the longer operon transcript, lytM cannot be translated because its ribosome-binding site is sequestered into a translationally inactive secondary structure. In addition, the effector of the agr system, RNAIII, can inhibit translation of lytM present on the operon without altering the transcript level but does not have an effect on the translation of the upstream gene. We propose that this dual regulation of lytM expression, at the transcriptional and post-transcriptional levels, contributes to prevent cell wall damage during the stationary phase of growth.

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Molecular cloning, sequencing, and expression of lytM, a unique autolytic gene of Staphylococcus aureus

Cited by 79 publications

References 33 publications

Identification of Genes Controlled by the Essential YycG/YycF Two-Component System ofStaphylococcus aureus

Identification of Genes Controlled by the Essential YycG/YycF Two-Component System ofStaphylococcus aureus

The amoebae plate test implicates a paralogue of lpxB in the interaction of Legionella pneumophila with Acanthamoeba castellanii

Various checkpoints prevent the synthesis ofStaphylococcus aureuspeptidoglycan hydrolase LytM in the stationary growth phase

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