Genetics of Treponema: relationship between Treponema pallidum and five cultivable treponemes

Miao, R M; Fieldsteel, A H

doi:10.1128/jb.133.1.101-107.1978

Cited by 72 publications

(47 citation statements)

References 14 publications

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“…The inability of T. phagedenis biotype Reiter proteins to significantly alter the T. paUidum immunoprecipitation gel profile (Fig. 8) verifies the specificity of the reaction and the fact that little antigenic homology exists between these treponemes, as previously suggested by DNA hybridization studies (28). However, further serologic analysis with an increased antigenic load is required to complete the analysis of antigenic cross-reactivity.…”

Section: Discussionsupporting

confidence: 75%

Molecular characterization of receptor binding proteins and immunogens of virulent Treponema pallidum.

Baseman¹,

Hayes²

1980

The Journal of Experimental Medicine

123

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Receptor binding proteins of Treponema pallidum were identified by incubation of [35S]methionine-labeled, soluble T. pallidum preparations with formaldehyde-fixed HEp-2 cells. Three major treponemal proteins (bands 1--3) that avidly bound to the eucaryotic cell surface were detected by sodium dodecylsulfate-polyacrylamide gel electrophoresis and fluorography. Brief trypsin treatment of HEp-2 cells before formaldehyde fixation reduced the extent of the interaction of these treponemal macromolecules, which implicated receptor-mediated attachment mechanisms. The presence of unlabeled T. pallidum preparations directly competed with radiolabeled T. pallidum samples for the available HEp-2 cells, which suggested a limiting number of membrane binding sites. Samples of unlabeled avirulent Reiter treponeme did not compete. T. Pallidum immunogens were examined by radioimmunoprecipitation with human and rabbit syphilitic sera. Of interest were the similarities and extent of the humoral response represented by the detection of antigen-antibody complexes against numberous treponemal proteins, including bands 1--3. T. pallidum portein band 1 appeared to be the major antigenic stimulus. Formation of antigen-antibody complexes between 35S-labeled T. pallidum proteins and human syphilitic sera was prevented by unlabeled T. pallidum but not by T. phagedenis preparations, which demonstrated specificity of the reaction. Gel profiles of radioimmunoprecipitation assays using radiolabeled T. pallidum antigens and human syphilitic and yaws sera delineated both the similarities and differences in the humoral response to these two spirochetes. The latter suggested both overlapping and distinguishing antigenic properties between T. pallidum and T. pertenue. Detection in yaws sera of specific antibody against T. pallidum protein bands 1--3 further incriminates the role of these three treponemal proteins as virulence determinants.

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Section: Discussionsupporting

confidence: 75%

Molecular characterization of receptor binding proteins and immunogens of virulent Treponema pallidum.

Baseman¹,

Hayes²

1980

The Journal of Experimental Medicine

123

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“…Unlike indirect methods of calculating genome molecular weight, PFGE allows visualization of large DNA fragments and measurement of the lengths of these fragments, thus providing a direct and accurate means of determining the physical sizes of bacterial genomes (11,17). Given the agreement between the genome size estimates obtained by digestion of T. pallidum DNA with NotI and SpeI, the T. pallidum genome size reported in this study appears accurate and corrects the previous size estimate (13). The reason for the high estimated size of the T. pallidum genome calculated from renaturation kinetics is not clear.…”

Section: Discussionsupporting

confidence: 66%

“…Previously, the genome ofT. pallidum was calculated on the basis of renaturation kinetics to be 13,700 kbp (9.05 x 109 Da [13]). Unlike indirect methods of calculating genome molecular weight, PFGE allows visualization of large DNA fragments and measurement of the lengths of these fragments, thus providing a direct and accurate means of determining the physical sizes of bacterial genomes (11,17).…”

Section: Discussionmentioning

confidence: 99%

Treponema pallidum subsp. pallidum has a single, circular chromosome with a size of approximately 900 kilobase pairs

Walker¹,

Arnett²,

Heath³

et al. 1991

Infect Immun

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The genome size and chromosome conformation of Treponema pallidum subsp. pallidum, Nichols strain, were determined by contour-clamped homogeneous electric field electrophoresis, a pulsed-field gel electrophoresis technique. Digestion of T. pallidum subsp. pallidum DNA with the restriction endonucleases NotI and SpeI produced 12 and 26 fragments, respectively. Summation of the physical lengths of the fragments produced by NotI and SpeI cleavage yielded average sizes of 900 and 913 kbp, respectively, for the genome of T. pallidum subsp. pallidum. Contour-clamped homogeneous electric field electrophoresis of T. pallidum subsp. pallidum DNA exposed to 4 krads of gamma irradiation resolved a single band of 800 to 1,000 kbp; treatment of the DNA with 16 krads of gamma irradiation resulted in the production of smaller fragments, whereas untreated DNA did not migrate into the gels. The gamma irradiation results indicate that T. pallidum subsp. pallidum has a single, circular chromosome that was linearized at a dosage of 4 krads of gamma irradiation. The size estimate provided by restriction endonuclease digestion with NotI and SpeI shows that the genome of T. pallidum subsp. pallidum, at approximately 900 kbp, is considerably smaller than the 13,700-kbp genome size calculated from renaturation kinetics.

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“…MGDG was easily detected by complement fixation reactions in T. phagedenis biotypes reiterii and phagedenis but not in T. pallidum (6). The absence of MGDG in T. pallidum is interesting in relation to evidence indicating the lack of genetic relatedness between T. pallidum and five cultivable Treponema (20). It would also be interesting to determine whether T. pertenue and T. carateum, treponemes pathogenic for humans which are morphologically and immunologically similar to T. pallidum, might resemble T. pallidum in lacking MGDG.…”

Section: Discussionmentioning

confidence: 99%

Unique lipid composition of Treponema pallidum (Nichols virulent strain)

Matthews¹,

Yang²,

Jenkin³

1979

Infect Immun

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The lipid composition of Treponema pallidum (Nichols virulent strain) was determined after purification of the organisms from the infected testes of corticosteroid-treated rabbits by differential centrifugation, filtration through Nuclepore membranes, and sedimentation in Hypaque density gradients. The total lipids were comprised of 32.2% neutral lipids, mainly cholesterol, and 67.8% phospholipids consisting of phosphatidylcholine (32.1%), sphingomyelin (14.8%), cardiolipin (13.0%), phosphatidylethanolamine (6.2%), phosphatidylinositol-serine (1.2%), and lysophosphatidylcholine (0.4%). Monoglycosyldiglyceride, a glycolipid comprising 25 to 50% of the total lipid of all Treponema previously examined, was not detected. The fatty acid composition was similar but quantitatively distinct from that of the infected testes tissue.Attempts to cultivate Treponema pallidum, the causative agent of syphilis, in vitro have failed since its discovery almost 75 years ago. Largely due to this, little is known about the organism's lipid metabolism or composition. Many cultivable Treponema require long-chain fatty acids for growth (9,16,22) since they can neither synthesize, 8-oxidize, nor desaturate fatty acids (1,10,19). Some intestinal and oral strains, however, can utilize short-chain fatty acids (26). Recent studies suggest that T. pallidum may also have lesions in lipid metabolism similar to many cultivable treponemes (9,18,21,25).Lipids comprise 15 to 20% of the cellular dry weight of the cultivable treponemes (10, 16). The major component is a glycolipid, monoglycosyldiglyceride (MGDG), containing galactose generally as the sugar moiety. MGDG comprises 25 to 50% of the total lipid of each of the approximately 20 treponemal strains of genital, oral, and intestinal origin so far examined (10,16,19) including T. hyodysenteriae (our unpublished data), the only cultivable pathogenic Treponema (12). Although Vaczi et al. (29) investigated the lipid composition of T. pallidum (Budapest strain), as well as four cultivable treponemes, they reported no quantitative data about the individual lipids or gave any indication of the glycolipid content of the organisms.Since T. pallidum cannot be cultivated in vitro, the organism is obtained for laboratory studies from the testes of experimentally infected rabbits. A major impedance of research on T. pallidum is the difficulty of obtaining large yields of the organism free of rabbit testicular material. In recent years, however, the use of corticosteroid-treated rabbits (8), Nuclepore filters (25), and Hypaque gradients (2) has proven remarkably useful in reducing this problem. We have used all of these tools to obtain high numbers of purified T. pallidum for lipid analysis. Infected rabbit testicular tissue and T. phagedenis biotype reiterii were also examined as comparative controls. Our results have shown that T. pallidum appears to be unique among Treponema in that the major lipid component of other Treponema, namely MGDG, was not detected in this human pathogen.MATERIALS AND METHODS C...

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Genetics of Treponema: relationship between Treponema pallidum and five cultivable treponemes

Cited by 72 publications

References 14 publications

Molecular characterization of receptor binding proteins and immunogens of virulent Treponema pallidum.

Molecular characterization of receptor binding proteins and immunogens of virulent Treponema pallidum.

Treponema pallidum subsp. pallidum has a single, circular chromosome with a size of approximately 900 kilobase pairs

Unique lipid composition of Treponema pallidum (Nichols virulent strain)

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