Identification and sequence analysis of a silent gene (ushA0) in Salmonella typhimurium

Burns, Dennis M.; Beacham, Ifor R.

doi:10.1016/0022-2836(86)90358-x

Cited by 32 publications

(32 citation statements)

References 55 publications

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“…Comparison of the NucA protein sequence from strain P860295 to that from the other strains showed a strikingly high degree of amino acid homology among all the strains sequenced. The NucA sequences from four strains (P880859, TN106, SH1015, and Eagan) are 100% identical, while those of the other five strains differ by 1 to 7 amino acid residues, as shown in Table 5 tidase protein sequences from rat (25), human (26), Vibrio parahaemolyticus NutA (44), and E. coli UshA (10). Two small regions of striking homology were observed among the 5Ј nucleotidase proteins as depicted in Fig.…”

Section: Immunogenicity Of Rnucamentioning

confidence: 92%

Identification of a Haemophilus influenzae 5′-Nucleotidase Protein: Cloning of the nucA Gene and Immunogenicity and Characterization of the NucA Protein

et al. 2000

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We report on the identification of a surface-exposed, highly conserved, immunogenic nontypeable Haemophilus influenzae (NTHi) protein, which elicits cross-reactive bactericidal antibodies against NTHi. The protein was extracted from NTHi strain P860295 with KSCN and purified; it migrated as a single band on a sodium dodecyl sulfate-polyacrylamide gel with an apparent molecular mass of 63 kDa. Mouse antiserum generated against the purified protein was reactive on whole-cell enzyme-linked immunosorbent assay (ELISA) with seven NTHi strains and type b Eagan and Whittier strains and exhibited bactericidal activity to homologous and heterologous NTHi strains. However, the protein is made in small amounts in NTHi as corroborated by immunoelectron microscopy. To further study this protein, we cloned, sequenced, and expressed it recombinantly in Escherichia coli. The recombinant protein is localized in the periplasm of E. coli and has been purified to homogeneity. Both the recombinant and native proteins possess 5-nucleotidase activity; hence, the protein has been called NucA. Mouse antiserum directed against the recombinant NucA protein was reactive on Western immunoblots and whole-cell ELISA with all H. influenzae strains tested including Eagan and was bactericidal for two heterologous strains tested. The antiserum also resulted in a log reduction in bacteremia, in an infant-rat protection study with H. influenzae type b as the challenge strain. These features suggest that NucA is a potential subunit vaccine candidate against NTHi disease.Nontypeable Haemophilus influenzae (NTHi) isolates are causative agents of a wide range of human diseases including tracheobronchitis, pneumonia, epiglottitis, and otitis media (7,28,47). Most vaccine development effort has been directed toward integral outer membrane components of NTHi. Outer membrane proteins (OMPs)

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Section: Immunogenicity Of Rnucamentioning

confidence: 92%

Identification of a Haemophilus influenzae 5′-Nucleotidase Protein: Cloning of the nucA Gene and Immunogenicity and Characterization of the NucA Protein

et al. 2000

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“…ebsC encodes a peptide of 164 amino acids that shows 36% identity to a 159-residue Salmonella typhimurium peptide (Fig. 5) that may be involved in the suppression of transcription of the silent gene ushA°, a nonexpressed gene homologous to the E. coli gene ushA, encoding a UDPsugar hydrolase (4). ebsB and ebsA translate to peptides of 135 and 153 amino acids, respectively.…”

Section: Junctionmentioning

confidence: 99%

Cloning and molecular analysis of genes affecting expression of binding substance, the recipient-encoded receptor(s) mediating mating aggregate formation in Enterococcus faecalis

Bensing

Dunny

1993

J Bacteriol

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Transfer of the conjugative plasmid pCF10 in Enterococcus faecalis strains involves production of a plasmid-encoded aggregation substance on the surface of donor cells in response to stimulation by a pheromone secreted by recipient cells. Aggregation substance then facilitates attachment to recipient cells via a chromosomally encoded receptor, termed binding substance (BS). A BS mutant, strain INY3000, generated by random Tn916 insertions, was previously found to carry copies of the transposon at four unique sites (K. M. Trotter and G. M. Dunny, Plasmid 24:57-67, 1990). In the present study, DNA flanking the Tn916 insertions was used to complement the BS mutation of INY3000 following Tn916 excision from cloned chromosomal fragments. Complementation results showed that three of the four regions mutated in INY3000 play some role in BS expression. Tn5 mutagenesis and DNA sequence analysis of the complementing fragment from one of these regions indicated the presence of three genes (ebsA, ebsB, and ebsC) that affect BS expression. The ebsA and ebsB genes encode peptides likely to function in cell wall metabolism, whereas ebsC may encode a product that suppresses the function or expression of EbsB.

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“…Phagemid-encoded proteins were selectively labelled using maxicells prepared from E. coli CSR603 and electrophoresed on 12% sodium dodecyl sulphate (SDS)^poly-acrylamide gels which were treated with`Amplify' (Amersham) and £uorographed [1,5].…”

Section: In Vivo Translation Using Maxicell Analysismentioning

confidence: 99%

“…In addition to these active ush alleles, we have identi¢ed two inactive alleles originally designated ushA 0 in S. enterica, and ushB 0 in E. coli [5,6]. The former allele, found in most natural isolates of S. enterica serovar Typhimurium, as well as in strains of serovars Gallinarum and Pullorum, is now referred to as ushA c , a newly de¢ned type of cryptic gene (see [7]), since an inactive 5P-nucleotidase is made [6,7].…”

Section: Introductionmentioning

confidence: 99%

The Escherichia coli orthologue of the Salmonella ushB gene (ushBc) produces neither UDP-sugar hydrolase activity nor detectable protein, but has an identical sequence to that of Escherichia coli cdh

Schroder

2001

FEMS Microbiology Letters

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The Escherichia coli orthologue of the Salmonella ushB gene (ushB c ) produces neither UDP-sugar hydrolase activity nor detectable protein, but has an identical sequence to that of Escherichia coli cdh Abstract Salmonella ushB, which encodes a membrane-bound UDP-sugar hydrolase, has an Escherichia coli orthologue (ushB c ) which does not detectably produce this activity. In this report, we show that ushB c does not produce any detectable protein either, despite being transcribed normally. Remarkably, ushB c is shown to have 100% sequence identity with E. coli cdh, previously characterised as encoding an active CDPdiglyceride hydrolase, an apparent contradiction with implications regarding enzyme evolution. We suggest that a useful gene designation is cdh (ushB c ) rather than either ushB

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Identification and sequence analysis of a silent gene (ushA0) in Salmonella typhimurium

Cited by 32 publications

References 55 publications

Identification of a Haemophilus influenzae 5′-Nucleotidase Protein: Cloning of the nucA Gene and Immunogenicity and Characterization of the NucA Protein

Identification of a Haemophilus influenzae 5′-Nucleotidase Protein: Cloning of the nucA Gene and Immunogenicity and Characterization of the NucA Protein

Cloning and molecular analysis of genes affecting expression of binding substance, the recipient-encoded receptor(s) mediating mating aggregate formation in Enterococcus faecalis

The Escherichia coli orthologue of the Salmonella ushB gene (ushBc) produces neither UDP-sugar hydrolase activity nor detectable protein, but has an identical sequence to that of Escherichia coli cdh

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