Pathogenic Mycoplasmas: Cultivation and Vertebrate Pathogenicity of a New Spiroplasma

Jg, Tully; Whitcomb, R. F.; Clark, H Fred; Dl, Williamson

doi:10.1126/science.841314

Cited by 370 publications

(183 citation statements)

References 18 publications

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“…The nomenclature indicates the site of transposon insertion by open reading frame (ORF) (MPN247 or MPN248), followed by the last amino acid residue encoded by that truncated ORF (15). Cultures were grown to mid-log phase in SP-4 medium (30), with the addition of 18 g/ml gentamicin and 24 g/ml chloramphenicol where appropriate for antibiotic selection. Hemadsorption and satellite growth were characterized as described previously (8).…”

Section: Methodsmentioning

confidence: 99%

Protein Kinase/Phosphatase Function Correlates with Gliding Motility in Mycoplasma pneumoniae

Page

Krause

2013

J Bacteriol

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Mycoplasma pneumoniae exhibits a novel form of gliding motility that is mediated by the terminal organelle, a differentiated polar structure. Given that genes known to be involved in gliding in other organisms are absent in M. pneumoniae, random transposon mutagenesis was employed to generate mutants with gliding-deficient phenotypes. Transposon insertions in the only annotated Ser/Thr protein kinase gene (prkC; MPN248) and its cognate phosphatase gene (prpC; MPN247) in M. pneumoniae resulted in significant and contrasting effects on gliding frequencies. prkC mutant cells glided at approximately half the frequency of wild-type cells, while prpC mutant cells glided more than twice as frequently as wild-type cells. Phosphoprotein staining confirmed the association between phosphorylation of the cytoskeletal proteins HMW1 and HMW2 and membrane protein P1 and the gliding phenotype. When the prpC mutant was complemented by transposon delivery of a wild-type copy of the prpC allele, gliding frequencies and phosphorylation levels returned to the wild-type standard. Surprisingly, delivery of the recombinant wild-type prkC allele dramatically increased gliding frequency to a level approximately 3-fold greater than that of wild-type in the prkC mutant. Collectively, these data suggest that PrkC and PrpC work in opposition in M. pneumoniae to influence gliding frequency.M ycoplasma pneumoniae is a cell wall-less bacterial pathogen of the human respiratory tract causing primary atypical pneumonia and tracheobronchitis (1). Mycoplasmas lack major biosynthetic pathways, classical transcriptional regulators, chemotactic and other two-component systems, and the prototypical prokaryotic cell division apparatus (2, 3). The limited biosynthetic capabilities of mycoplasmas are generally explained by their evolution as obligate parasites of diverse eukaryotic hosts (4). Colonization of the host respiratory epithelium by M. pneumoniae requires gliding motility (5), which might facilitate access to receptors on the host cell surface and subsequent lateral spread.The gliding apparatus of M. pneumoniae is a polar terminal structure (6) that also functions in cell division (7) and adhesion to host receptors (2, 5). While the cytoskeletal protein HMW1 (MPN447) and membrane proteins P1, B/C, and P30 (MPN141, MPN142, and MPN453, respectively) localize to the terminal organelle and are required for gliding (5,(8)(9)(10), these proteins are also essential for cytadherence and attachment to surfaces. Accordingly, their distinct functions in gliding motility are difficult to define by mutagenesis alone. Given that the M. pneumoniae genome exhibits no homology to elements of defined gliding mechanisms (2, 3), including those of other gliding mycoplasmas (11-14), it was necessary to perform saturating transposon mutagenesis in order to identify the components specific to gliding (15). Transposon insertions in the genes encoding the cytoskeletal proteins P41 and P65 (MPN311 and MPN309, respectively) (16), which are known to localize to the termina...

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Section: Methodsmentioning

confidence: 99%

Protein Kinase/Phosphatase Function Correlates with Gliding Motility in Mycoplasma pneumoniae

Page

Krause

2013

J Bacteriol

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“…The remainder was centrifuged again for 10 min at the same speed at 4"C, and the supernatant was then forced, by hand pressure, through a membrane filter (pore size, 220 nm). Tubes containing 1 ml of SP-4 medium (35,36) were inoculated with 0.3 ml of filtrate and incubated at 30°C. After 10 days, the pH indicator in the medium had shifted to the acid color, indicating that growth had occurred.…”

Section: Methodsmentioning

confidence: 99%

Spiroplasma sabaudiense sp. nov. from Mosquitoes Collected in France

Abalain-Colloc¹,

Chastel²,

Tully

et al. 1987

International Journal of Systematic Bacteriology

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“…hominis ATCC 231 14, Acholeplasma laidlawii ATCC 23206 and Ureaplasma urealyticum ATCC 27618. All mycoplasma strains were cultured in diphasic SP-4 medium (Tully et al, 1977) containing 2000 units penicillin G ml-', 500 units polymyxin B ml-' and 2.5 pg amphotericin B ml-', as described by . Thallium acetate was not added to the medium since it can inhibit growth of M .…”

Section: Methodsmentioning

confidence: 99%

Use of a triplex polymerase chain reaction for the detection and differentiation of Mycoplasma pneumoniae and Mycoplasma genitalium in the presence of human DNA

Cadieux

Lebel²,

Brousseau³

1993

Journal of General Microbiology

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PCR primers corresponding to the adhesin genes of Mycoplasma pneumoniae and Mycoplasma genitalium were shown to detect the corresponding organisms specifically. Absence of cross-reaction with seven other mollicute species and six unrelated bacterial species commonly found in humans was demonstrated. Positive control primers directed against human mitochondria1 DNA could be mixed with the Mycoplasma primers without loss of specificity or sensitivity. A detection level of 10 c.f.u. of either Mycoplasma species could be readily obtained, even in the presence of lo4 human cells. The triplex PCR method developed is very simple and does not require hybridization or the use of radioisotopes and allows detection and differentiation of these mycoplasmas against the background of human DNA found in clinical specimens.

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Pathogenic Mycoplasmas: Cultivation and Vertebrate Pathogenicity of a New Spiroplasma

Cited by 370 publications

References 18 publications

Protein Kinase/Phosphatase Function Correlates with Gliding Motility in Mycoplasma pneumoniae

Protein Kinase/Phosphatase Function Correlates with Gliding Motility in Mycoplasma pneumoniae

Spiroplasma sabaudiense sp. nov. from Mosquitoes Collected in France

Use of a triplex polymerase chain reaction for the detection and differentiation of Mycoplasma pneumoniae and Mycoplasma genitalium in the presence of human DNA

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