Cloning and nucleotide sequence of the M1-encoding gene from Streptomyces globisporus

Lichenstein, Henri S.; Hastings, Alice; Langley, Keith; Mendiaz, Elizabeth A.; Rohde, Michael F.; Elmore, R.H.; Zukowski, Mark M.

doi:10.1016/0378-1119(90)90062-v

Cited by 39 publications

(19 citation statements)

References 25 publications

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“…Looking for a signature that would point to the catalytic site of adh lysin, we compared its sequence with the N terminus of lysozyme Ch of the fungus Chalaropsis sp. (19), which was previously shown to be similar to the N termini of muramidase CPL-1 of S. pneumoniae phage Cp-1 (24), N-acetylmuramidase M1 of Streptomyces globisporus (47), and LysA of L. bulgaricus phage mv1 (8). Lysozyme Ch, CPL-1, and M1 all contain an Asp and a Glu residue 26 or 27 amino acids apart, which, in the case of the Chaloropsis enzyme, were shown to be involved in catalytic activity (22).…”

Section: Discussionmentioning

confidence: 75%

Primary structure and functional analysis of the lysis genes of Lactobacillus gasseri bacteriophage phi adh

1995

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The lysis genes of the Lactobacillus gasseri bacteriophage adh were isolated by complementation of a lambda Sam mutation in Escherichia coli. Nucleotide sequencing of a 1,735-bp DNA fragment revealed two adjacent coding regions of 342 bp (hol) and 951 bp (lys) in the same reading frame which appear to belong to a common transcriptional unit. Proteins corresponding to the predicted gene products, holin (12.9 kDa) and lysin (34.7 kDa), were identified by in vitro and in vivo expression of the cloned genes. The adh holin is a membrane-bound protein with structural similarity to lysis proteins of other phage, known to be required for the transit of murein hydrolases through the cytoplasmic membrane. The adh lysin shows homology with mureinolytic enzymes encoded by the Lactobacillus bulgaricus phage mv4, the Streptococcus pneumoniae phage Cp-1, Cp-7, and Cp-9, and the Lactococcus lactis phage LC3. Significant homology with the N termini of known muramidases suggests that adh lysin acts by a similar catalytic mechanism. In E. coli, the adh lysin seems to be associated with the total membrane fraction, from which it can be extracted with lauryl sarcosinate. Either one of the adh lysis proteins provoked lysis of E. coli when expressed along with holins or lysins of phage lambda or Bacillus subtilis phage 29. Concomitant expression of the combined holin and lysin functions of adh in E. coli, however, did not result in efficient cell lysis.Large Escherichia coli phage in general appear to encode at least two lysis functions, a murein hydrolase, required for destruction of the peptidoglycan, and a protein termed holin which permits access of the lytic enzyme to the periplasm (for a review, see reference 78). In the case of bacteriophage lambda, oligomerization of the holin (protein S) in the inner membrane of E. coli apparently leads to formation of a nonspecific lesion through which the lambda transglycosylase is released to the periplasm at the end of the vegetative cycle (80). The expression of the S gene and thus the kinetics of formation of the S-dependent hole in the inner membrane is tightly controlled at the transcriptional (35) and at the translational level (7, 53), as well as posttranslationally by virtue of two S-encoded polypeptides with opposing functions (6, 70). Holin functions have also been attributed to the products of P22 gene 13 (58), to phage 21 gene S (9), and recently to protein 14 of the Bacillus subtilis phage 29 (70), the first holin identified from a phage of gram-positive bacteria. Large phage therefore appear to pursue an evolutionarily conserved lysis pathway.In phage lambda, P22, 21, and 29, the genes encoding the corresponding holins and murein hydrolases are arranged identically. The holin gene in all cases precedes the gene encoding the murein hydrolase and overlaps at least with its ribosome-binding site (9,13,30,58). With the exception of the products of the lambda S and P22 13 genes, which are nearly identical (58), the known holins show no homology with each other (78). Since lambda S...

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Section: Discussionmentioning

confidence: 75%

Primary structure and functional analysis of the lysis genes of Lactobacillus gasseri bacteriophage phi adh

1995

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“…Distinct classes of muramidases have been discerned based on amino acid sequence similarities and specific functional properties (17,18). It has been suggested that the muramidases of the fungus Chalaropsis (5), the bacterium Streptomyces globisporous (18), the bacterium Clostridium acetobutylicum (4), and the pneumococcal bacteriophages of the Cp family (11) form a separate group within the lysozyme family (4,18).…”

Section: Discussionmentioning

confidence: 99%

“…It has been suggested that the muramidases of the fungus Chalaropsis (5), the bacterium Streptomyces globisporous (18), the bacterium Clostridium acetobutylicum (4), and the pneumococcal bacteriophages of the Cp family (11) form a separate group within the lysozyme family (4,18). In the Chalaropsis-type muramidases, the similarity observed among the enzymes corresponds to the amino-terminal half of the molecules, and the two amino acids of the muramidase of Chalaropsis, Asp-6 and Glu-33, that have been reported to be involved in the active center of the enzyme (9) have also been found in those enzymes, where they are conserved in equivalent positions.…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization of a germination-specific muramidase from Clostridium perfringens S40 spores and nucleotide sequence of the corresponding gene

et al. 1997

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The exudate of fully germinated spores of Clostridium perfringens S40 in 0.15 M KCl-50 mM potassium phosphate (pH 7.0) was found to contain another spore-lytic enzyme in addition to the germination-specific amidase previously characterized (S. Miyata, R. Moriyama, N. Miyahara, and S. Makino, Microbiology 141:2643-2650, 1995). The lytic enzyme was purified to homogeneity by anion-exchange chromatography and shown to be a muramidase which requires divalent cations (Ca ) for its activity. The enzyme was inactivated by sulfhydryl reagents, and sodium thioglycolate reversed the inactivation by Hg 2؉. The muramidase hydrolyzed isolated spore cortical fragments from a variety of wild-type organisms but had minimal activity on decoated spores and isolated cell walls. However, the enzyme was not capable of digesting isolated cortical fragments from spores of Bacillus subtilis ADD1, which lacks muramic acid ␦-lactam in its cortical peptidoglycan. This indicates that the enzyme recognizes the ␦-lactam residue peculiar to spore peptidoglycan, suggesting an involvement of the enzyme in spore germination. Immunochemical studies indicated that the muramidase in its mature form is localized on the exterior of the cortex layer in the dormant spore. A gene encoding the muramidase, sleM, was cloned into Escherichia coli, and the nucleotide sequence was determined. The gene encoded a protein of 321 amino acids with a deduced molecular weight of 36,358. The deduced amino acid sequence of the sleM gene indicated that the enzyme is produced in a mature form. It was suggested that the muramidase belongs to a separate group within the lysozyme family typified by the fungus Chalaropsis lysozyme. A possible mechanism for cortex degradation in C. perfringens S40 spores is discussed.Bacterial spore germination, defined as the irreversible loss of spore characteristics, is triggered by specific germinants and proceeds through a set of sequential steps. The later stage of germination involves decomposition of the spore peptidoglycan by a spore cortex-lytic enzyme(s) (8,24). The enzymes involved in the reaction have been identified from spores of Bacillus megaterium (7), Bacillus cereus (20, 26), Bacillus subtilis (25), and Clostridium perfringens (22, 23) and characterized as germination-specific amidases. A 30-kDa amidase of B. subtilis contributes to germination of the organism mediated by Lalanine but not to germination triggered by an unspecified germinant(s) present in culture medium such as Schaffer's medium (25). This suggests the presence of multiple pathways for spore germination, and it is likely that a germination-specific enzyme(s) other than the amidases identified to date is also involved in hydrolysis of spore peptidoglycan.Gombas and Labbe reported (14) that spore cortex-lytic enzymes differing in their substrate specificity are released during germination of C. perfringens spores, though it was not clear whether the enzymes are actually involved in germination; one such enzyme lyses decoated spores, and the other hydrolyzes i...

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“…1 shows the HPLC elution profiles of muropeptide species generated from the peptidoglycans of the parental strain COL and its vancomycin-resistant (VM) and teicoplanin-resistant (TNM) derivatives, and a third mutant, TM, selected from mutant VM as a highly teicoplanin-resistant derivative (6). The HPLC profiles in panels A show muropeptide species obtained after treatment of the peptidoglycans with the M1 muramidase, an enzyme that breaks glycosidic bonds between the disaccharide units in the peptidoglycan (14). Drastic reduction in the proportion of highly cross-linked muropeptide oligomers (i.e.…”

Section: Methodsmentioning

confidence: 99%

Inactivated pbp4 in Highly Glycopeptide-resistant Laboratory Mutants of Staphylococcus aureus

Sieradzki

Pinho

Tomasz

1999

Journal of Biological Chemistry

125

110

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Microb. Drug Resist. 4, 159 -168). We now report that the distorted peptidoglycan composition is related to defects in penicillin-binding protein 4 (PBP4); no PBP4 was detectable by the fluorographic assay in membrane preparations from the mutants, and comparison of the sequence of pbp4 amplified from the mutants indicated disruption of the gene by two types of abnormalities, a 17-amino acid long duplication starting at position 305 of the pbp4 gene was detected in the vancomycin-resistant mutant, and a stop codon was found to be introduced into the pbp4 KTG motif at position 261 in the mutant selected for teicoplanin resistance. Additional common patterns of disturbances in the peptidoglycan metabolism of the mutants are indicated by the increased sensitivity of mutant cell walls to the M1 muramidase and decreased sensitivity to lysostaphin, which is a reversal of the susceptibility pattern of the parental cell walls. Furthermore, the results of high performance liquid chromatography analysis of lysostaphin digests of peptidoglycan suggest an increase in the average chain length of the glycan strands in the peptidoglycan of the glycopeptide-resistant mutants. The increased molar proportion of muropeptide monomers in the cell wall of the glycopeptide-resistant mutants should provide binding sites for the "capture" of vancomycin and teicoplanin molecules, which may be part of the mechanism of glycopeptide resistance in S. aureus.

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Cloning and nucleotide sequence of the M1-encoding gene from Streptomyces globisporus

Cited by 39 publications

References 25 publications

Primary structure and functional analysis of the lysis genes of Lactobacillus gasseri bacteriophage phi adh

Primary structure and functional analysis of the lysis genes of Lactobacillus gasseri bacteriophage phi adh

Molecular characterization of a germination-specific muramidase from Clostridium perfringens S40 spores and nucleotide sequence of the corresponding gene

Inactivated pbp4 in Highly Glycopeptide-resistant Laboratory Mutants of Staphylococcus aureus

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