Molecular cloning and genetic characterization of the rfb region from Yersinia pseudotuberculosis serogroup IIA, which determines the formation of the 3,6-dideoxyhexose abequose

Kessler, Annette C.; Brown, Peter K.; Romana, Lajwant K.; Reeves, Peter R.

doi:10.1099/00221287-137-12-2689

Cited by 26 publications

(18 citation statements)

References 22 publications

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“…Initiation towards this goal began with the cloning and analysis of the Salmonella typhimurium rfb (0-antigen) cluster, which led to the elucidation of the entire sequence of this region and the identification of 12 of the 15 genes present in the cluster (6,15,46). Subsequently, the deoxy and dideoxy genes within the rfb clusters of S. typhimurium (group B, which contains abequose) were compared to those within Salmonella paratyphi (group A, which contains paratose), Salmonella typhi (group D, which contains tyvelose), and, most recently, Y pseudotuberculosis M85 (serogroup IIA, which contains abequose) (17,18). As shown in Fig.…”

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confidence: 99%

Studies of the biosynthesis of 3,6-dideoxyhexoses: molecular cloning and characterization of the asc (ascarylose) region from Yersinia pseudotuberculosis serogroup VA

et al. 1994

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The 3,6-dideoxyhexoses are found in the lipopolysaccharides of gram-negative bacteria, where they have been shown to be the dominant antigenic determinants. Of the five 3,6-dideoxyhexoses known to A large degree of bacterial immunological diversity in gram-negative species is attributed to the portion of the lipopolysaccharides known as the 0 antigen. Exposed on the cell envelope, the 0 antigen is the dominant surface entity which is polymorphic and provides the basis for the serological classification of the family Enterobacteriaceae (4,16,26). Among the large number of monosaccharides found as components of 0-specific polysaccharides, derivatives of the 3,6-dideoxyhexoses have drawn special attention because of their highly immunogenic characteristics (22,43,45). It has been shown that only a limited number of species of gram-negative bacteria, all in the family Enterobacteraceae, are able to produce these unusual sugars, and, of the eight possible stereoisomers of 3,6-dideoxyhexoses, only five have been identified in nature. Although colitose (3, and abequose (3,6-dideoxy-D-ylo-hexose, compound I [see Fig. 1]) can be detected in the lipopolysaccharide of some Escherichia coli serotypes and Citrobacter strains, respectively, the five 3,6-dideoxyhexoses known to occur naturally are present mainly in Salmonella and Yersinia spp. as major antigenic determinants (23). For example, abequose, colitose, tyvelose (3,6-dideoxy-D-arabino-hexose, compound II), and paratose (3,6-dideoxy-D-ribo-hexose, compound III) have been found in strains of Salmonella enterica (34), while all five, including ascarylose (3,6-dideOxy-L-arabino-hexose, compound * Corresponding author. Phone: (612) 626-7541. t Present address:

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Section: Discussionmentioning

confidence: 99%

Studies of the biosynthesis of 3,6-dideoxyhexoses: molecular cloning and characterization of the asc (ascarylose) region from Yersinia pseudotuberculosis serogroup VA

et al. 1994

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“…In Salmonella serogroups A, B and D, the $ 6 gene is located outside the r + region (Collins & Hackett, 1991), while in Salmonella enterica serogroups C1, C2 and El, Sbigella hsenteriae type I, and SbigellafEexneri, $ 6 is located within the r -gene cluster (Brown et a/., 1991 ;Lee e t al., 1992;Wang e t al., 1992;Morona e t al., 1994). In the $6-independent polymerization represented by Escbericbia coli serotypes 0 8 and 0 9 , the 0-antigen is synthesized by processive transfer of sugars to a growing chain attached to undecaprenol pyrophosphate (Jann & Jann, 1984;Whitfield & Valvano, 1993).…”

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The gene cluster directing O-antigen biosynthesis in Yersinia enterocolitica serotype O:8: identification of the genes for mannose and galactose biosynthesis and the gene for the O-antigen polymerase

1996

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The rfb gene cluster of Yersinia entemcolitica serotype 0:8 (Ye08) strain 8081 -c was cloned by cosmid cloning. Restriction mapping, deletion analysis and transposon mutagenesis showed that about 19 kb of the cloned DNA is Kiinamyllynkatu 13, 20520 Turku, Finland essential for the synthesis and expression of the Ye08 0-side-chain in Escherichia coli. Deletion analysis generated a derivative that expressed semirough LPS, a phenotype typical of an rfc mutant lacking the 0-antigen polymerase. The deletions and transcomplementation experiments allowed localization of the rfc gene to the 3'-end of the rfb gene cluster. The deduced Ye08 Rfc did not share significant amino acid sequence similarity with any other protein, but its amino acid composition and hydrophobicity profile are similar to those of identified Rfc proteins. In addition, the codon usage of the rfc gene is similar to other rfc genes. Nucleotide sequence analysis identified three other genes upstream of rfc. Two of the gene products showed 60-70°/0 identity to the RfbM and RfbK proteins that are biosynthetic enzymes for the GDPmannose pathway of enterobacteria. The third gene product was about 50-80 O/ o identical to the bacterial GalE protein, UDPglucose bepimerase, which catalyses the epimerization of UDPglucose to UDPgalactose. Since mannose and galactose are both present in the Ye08 0-antigen repeat unit, the above three genes are likely to belong to the rib gene cluster. A gene similar to the gsk gene downstream of rfc, and genes similar to adk and hemH upstream of the rfb gene cluster, were recognized. Thus the gene cluster of Ye08 is located between the ad&-hemH and gsk loci, and the order is adk-hemH-dbfc-gsk in the chromosome. Also in other Yersinia spp., the locus downstream of the hemH gene is occupied by gene clusters associated with LPS biosynthesis.

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“…We have previously reported the cloning of the Y pseudotuberculosis serogroup IIA rib region (19). Hybridization studies of this abequose-producing strain (M85) had shown that at least some of the M85 rib genes are related to DDH pathway genes (rfbF and rCbG) of S. enterica LT2.…”

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“…We have previously reported the cloning of the Y pseudotuberculosis serogroup IIA rib region (19 (51).…”

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“…DNA regions outside rib are indicated in black. Previous mutagenesis data indicated that the left-hand end of the M85 rib region was approximately between kilobase positions 35 and 37 of the original cosmid clone pPR981 (19), which corresponds to kilobase positions 0 to 2 in the DNA sequence. Sequence analysis showed extensive noncoding DNA sequences upstream of orf0.8, indicating that the M85 rib cluster, in fact, begins with orfO.8.…”

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Molecular analysis of the 3,6-dideoxyhexose pathway genes of Yersinia pseudotuberculosis serogroup IIA

1993

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-B, and -C; IVA and -B; and VA and -B), is due to differences in sugar composition and arrangement of the remaining 0 unit (35).In both S. enterica and Y pseudotuberculosis, DDH are formed by the same biosynthetic-reaction sequences. Four DDH, abequose, tyvelose, paratose, and ascarylose, are synthesized by a common pathway proceeding from CDP-D-glucose ( Fig. 1) (28); the fifth, colitose, is derived from GDP-D-mannose presumably via a similar reaction sequence (11). All enzymes involved in DDH biosynthesis in S. enterica are encoded within the rib gene cluster, which is * Corresponding author. responsible for the formation of the O-specific subunit of the LPS (26).The rfb gene clusters of several DDH-containing S. enterica strains have been cloned and sequenced (7,8,17,24,45,50), and most of the DDH-related genes have been identified. In all cases investigated, the arrangement of the DDH pathway genes and their relative positions within the rib region are conserved: immediately downstream of the rhamnose pathway genes, a block of four highly conserved genes is found, comprising rfbF and rfbG as well as two open reading frames (ORFs), orf7.6 and orflO.4, which are thought to correspond to the postulated DDH pathway genes rflI and OJbH (8). Abequose-forming strains of serogroups B and C2 (serovars typhimurium and muenchen, respectively) were shown to possess another gene, rfbJ, coding for abequose synthase, which is located adjacent to this highly conserved block; although secondary structure predictions indicated very similar proteins, DNA and amino acid sequences of the two rfbl genes had only low levels of similarity (only 36% identity at the amino acid level [8]). In strains forming tyvelose and paratose (serovars typhi and paratyphi of serogroups D and A, respectively), the rfbJ gene was replaced by a paratose synthase gene, rUbS, and a tyvelose epimerase gene, rfbE (46). The tyvelose epimerase gene in the paratose-producing group A strain, however, was found to be inactive because of a single point mutation.While rfbS still showed a low degree of similarity to rfib at the DNA sequence level, no counterpart to rfbE was found in the rfb region of the abequose-producing strains of S. enterica. Sequence analysis revealed unusually low G+C contents for all rib regions investigated, suggesting a relatively recent transfer of the gene cluster to S. enterica from a nonenterobacterial donor with a low G+C content (8,17,22,48).We have previously reported the cloning of the Y pseudotuberculosis serogroup IIA rib region (19). Hybridization studies of this abequose-producing strain (M85) had shown that at least some of the M85 rib genes are related to DDH pathway genes (rfbF and rCbG) of S. enterica LT2. In this study, we present the sequence and detailed analysis of the DDH pathway gene region of this Y pseudotuberculosis strain. Most of its DDH pathway genes were clearly related

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Molecular cloning and genetic characterization of the rfb region from Yersinia pseudotuberculosis serogroup IIA, which determines the formation of the 3,6-dideoxyhexose abequose

Cited by 26 publications

References 22 publications

Studies of the biosynthesis of 3,6-dideoxyhexoses: molecular cloning and characterization of the asc (ascarylose) region from Yersinia pseudotuberculosis serogroup VA

Studies of the biosynthesis of 3,6-dideoxyhexoses: molecular cloning and characterization of the asc (ascarylose) region from Yersinia pseudotuberculosis serogroup VA

The gene cluster directing O-antigen biosynthesis in Yersinia enterocolitica serotype O:8: identification of the genes for mannose and galactose biosynthesis and the gene for the O-antigen polymerase

Molecular analysis of the 3,6-dideoxyhexose pathway genes of Yersinia pseudotuberculosis serogroup IIA

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