Simultaneous Production of Two Capsular Polysaccharides by Pneumococcus

Austrian, Robert; Bernheimer, Harriet P.; Smith, Evelyn E. B.; Mills, George T.

doi:10.1084/jem.110.4.585

Cited by 69 publications

(25 citation statements)

References 17 publications

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“…In a study of the UPPG dehydrogenase activity of Streptococcus pneumoniae strain R19, derived from capsular type II, Smith et al (1959) observed the marked instability of this enzyme and the low yields of UPPGA obtained with the experimental conditions used. It was considered possible that the low dehydrogenase activity might be a function of inactivation of the enzyme activity by hydrogen peroxide (H,OJ.…”

Section: Resultsmentioning

confidence: 99%

Uridine Pyrophosphoglucose Dehydrogenase in Capsulated and Non-capsulated Strains of Pneumococcus Type I

Smith

Mills

Austrian

et al. 1960

Journal of General Microbiology

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SUMMARY:A method is presented for the estimation of uridine pyrophosphogluc o~e dehydrogenase mtivity in extracts of pneumococci; this utilizes 14C-labelled ddine pyrophosphoglucose (UPPG). This method overcomes the difBcu.Ity of the speotrophotom&ric assay caused by the presence of reduced diphosphopyridine nudeotide (DPNH) oKtdase in such extracts. An investigation was made o f ' the protective effof catalase and thioglycollate on pneumocoocal UPPG dehydrogems&. Both these substances prevented inactivation of this enzyme. The results suggest that inactivation is caused by hydrogen peroxide and that the point of attack on the exmyme is upon sulphydryl groups. The UPPG dehydrogenase activity of some capsulated and non-capsulated strains of type I pneumoeoccus was d e t e d e d . The capsulated and some of the non-capsulated strains examined had considerable UPPG dehydrogenase activity except one strain which had negligible activity. !Che enzyme UPPglucuronic acid-rh-epimerase from a type I pneumowwus requfred diphosphopyridine nucleotide for activity.

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Section: Resultsmentioning

confidence: 99%

Uridine Pyrophosphoglucose Dehydrogenase in Capsulated and Non-capsulated Strains of Pneumococcus Type I

Smith

Mills

Austrian

et al. 1960

Journal of General Microbiology

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“…The essential role of the capsule in S. pneumoniae virulence was established through the study of spontaneous mutants, the genetic transfer of capsular serotypes, and the use of a type 3-specific depolymerase to remove the capsule prior to infection (6,22,29). Genetic transfer of capsular serotypes indicated linkage of the genes necessary for synthesizing a specific capsular polysaccharide, and recombination experiments confirmed those linkages (4,18,37,38). A role for unlinked genes was indicated by the transfer of the normal capsule phenotype from mutants that produced reduced levels of capsule (28).…”

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confidence: 99%

Essential Role for Cellular Phosphoglucomutase in Virulence of Type 3 Streptococcus pneumoniae

et al. 2001

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Synthesis of the Streptococcus pneumoniae type 3 capsule requires the pathway glucose-6-phosphate (Glc-6-P) 3 Glc-1-P 3 UDP-Glc 3 UDP-glucuronic acid (UDP-GlcUA) 3 (GlcUA-Glc) n . The UDP-Glc dehydrogenase and synthase necessary for the latter two steps, and essential for capsule production, are encoded by genes (cps3D and cps3S, respectively) located in the type 3 capsule locus. The phosphoglucomutase (PGM) and Glc-1-P uridylyltransferase activities necessary for the first two steps are derived largely through the actions of cellular enzymes. Homologues of these enzymes, encoded by cps3M and cps3U in the type 3 locus, are not required for capsule production. Here, we show that cps3M and cps3U also are not required for mouse virulence. In contrast, nonencapsulated isolates containing defined mutations in cps3D and cps3S were avirulent, as were reduced-capsule isolates containing mutations in pgm. Insertion mutants that lacked PGM activity were avirulent in both immunologically normal (BALB/cByJ) and immunodeficient (CBA/N) mice. In contrast, a mutant (JY1060) with reduced PGM activity was avirulent in the former but had only modestly reduced virulence in the latter. The high virulence in CBA/N mice was not due to the lack of antibodies to phosphocholine but reflected a growth environment distinct from that found in BALB/cByJ mice. The reduced PGM activity of JY1060 resulted in enhanced binding of complement and antibodies to surface antigens. However, decomplementation of BALB/cByJ mice did not enhance the virulence of this mutant. Suppressor mutations, only some of which resulted in increased capsule production, increased the virulence of JY1060 in BALB/cByJ mice. The results suggest that PGM plays a critical role in pneumococcal virulence by affecting multiple cellular pathways.

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“…Nomendaure of Pnzumococcal Varients.--The nomenclature of pneumococcal variants employed will be that described in earlier publications (3)(4)(5). Mutants of noncapsulated variants producing capsules of C-like polysaccharide will be designated by the symbols denoting the parental noncapsulated strain followed by the symbol Cs; i.e., S-nC, is a variant of a noncapsulated strain of pneumoeoccns Type II which produces a capsule of soluble Clike polysaccharide.…”

Section: Methodsmentioning

confidence: 99%

“…It is known from earlier experiments that strains S-m4 and S-ms both possess the capsular genome of pneumococcus Type HI with mutations at different sites in the cistron controlling the synthesis of uridine diphosphoglucose dehydrogenase (4). When either strain is transformed with the DNA of the other, genetic recombination leads to correction of the defect in the synthesis of Type III capsular polysaccharide and fully capsulated Type III pneumococci are recovered.…”

Section: Phagocytosis Of C8 Pneumococci By Human Polymorpkonuclear Lementioning

confidence: 99%

“…When either strain is transformed with the DNA of the other, genetic recombination leads to correction of the defect in the synthesis of Type III capsular polysaccharide and fully capsulated Type III pneumococci are recovered. Additional studies of the genes controlling normal capsular synthesis have shown that they occupy the same region of the pneumococcal chromosome regardless of capsular type (4). In the present experiment, in which transformation of strain S-ms with DNA of strain S-m4C, led to the recovery of both capsulated Type IlI cells and C, cells, it is apparent that the genetic locus responsible for the production of C, capsular polysaccharide must occupy a site other than that controlling the synthesis of other pneumococcal capsular polysaccharides.…”

Section: Phagocytosis Of C8 Pneumococci By Human Polymorpkonuclear Lementioning

confidence: 99%

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CAPSULATION OF PNEUMOCOCCUS WITH SOLUBLE C-LIKE (Cs) POLYSACCHARIDE

Bornstein

Schiffman

Bernheimer

et al. 1968

The Journal of Experimental Medicine

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In 1949, Taylor (1) reported the recovery from a transformation reaction of a pneumococcus with a capsular antigen which differed both from that of the parent of the noncapsulated Type II strain which had served as the recipient cell and from that of the capsulated Type III strain which had served as the donor of deoxyribonucleates (DNA) in the system. In 1953, Beiser and Hotchldss (2) described the isolation of several analogous strains of capsulated pneumococci from a transforming system employing the same reactants. Antisera prepared in rabbits with vaccines of these strains gave a positive capsular precipitin or QueUung reaction with homologous cells and with other capsulated variants recovered from the same transforming system which failed to react with Type HI or Type II anticapsular serum. DNA from these unusual strains was shown also to be capable of transforming noncapsulated cells to the new capsular type. Further study of the novel capsular antigen failed to demonstrate a relationship between it and any of the 82 known pneumococcal capsular types. In subsequent experiments in our laboratories, a similar capsulated strain was recovered from a transforming reaction in which the recipient cells were noncapsulated variants of pneumococcus Type H; but, on this occasion, the DNA employed was extracted from capsulated Type I pneumococcal cells. Two hypotheses might account for the observations described. Uptake of foreign DNA might lead to recombination with or to rearrangement of resident DNA leading to the appearance of the novel capsular material; or the cells of the unusual capsular type might be spontaneous mutants selected during growth of cells in the transforming system. The experiments to be reported support the latter hypothesis and suggest that the novel capsule, composed of a C-like polysaccharide, may arise as a result of alteration of the normal mechanisms controlling the production of pneumococcal C or cell wall polysaccharide.

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Simultaneous Production of Two Capsular Polysaccharides by Pneumococcus

Cited by 69 publications

References 17 publications

Uridine Pyrophosphoglucose Dehydrogenase in Capsulated and Non-capsulated Strains of Pneumococcus Type I

Uridine Pyrophosphoglucose Dehydrogenase in Capsulated and Non-capsulated Strains of Pneumococcus Type I

Essential Role for Cellular Phosphoglucomutase in Virulence of Type 3 Streptococcus pneumoniae

CAPSULATION OF PNEUMOCOCCUS WITH SOLUBLE C-LIKE (Cs) POLYSACCHARIDE

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