A <i>Mycoplasma</i> genetic element resembling prokaryotic insertion sequences

Ferrell, Rebecca V; Heidari, M. B.; Wise, Kim S.; McIntosh, Mark A.

doi:10.1111/j.1365-2958.1989.tb00245.x

Cited by 36 publications

(20 citation statements)

References 49 publications

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“…Such mutations are known to occur both in the streptococcal M proteins (19,26) and in periodic coding regions (unrelated to Vlps) found in insertion sequence-like elements of M. hyorhinis and other swine mycoplasmas (13).…”

Section: Discussionmentioning

confidence: 99%

The Vlp system of Mycoplasma hyorhinis: combinatorial expression of distinct size variant lipoproteins generating high-frequency surface antigenic variation

1991

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Isogenic populations of Mycoplasma hyorhinis undergo in vitro high-frequency phase variation in the expression of surface lipoproteins; these products also vary markedly in size through changes in periodic protein structure (R. Rosengarten and K.S. Wise, Science 247:315-318, 1990). In this report, we rigorously define three distinct translation products comprising the Vlp (variable lipoprotein) system of M. hyorhinis SK76 and establish parameters of Vlp structural diversity and expression that distinguish the Vlp system from previously described examples of antigenic variation. VlpA, VlpB, and VlpC are prominent amphiphilic membrane lipoproteins characterized by detergent-phase fractionation and metabolic labeling with [35S]cysteine and [3H]palmitate. VlpA is distinguished from VlpB and VlpC by its selective labeling with [35S]methionine; VlpB and VlpC are distinguished by specific epitopes defined by surface-binding monoclonal antibodies (MAbs); a third MAb defines a surface epitope shared by VlpB and VlpC (but absent from VlpA). Each Vlp displays 12 to 30 spontaneous size variant forms comprising a periodic ladder that could also be generated by partial trypsin digestion of individual Vlp size variants. Different periodic intervals within VlpB and VlpC further distinguish these two products structurally. Mycoplasma colony opacity correlates inversely with Vlp size. Each Vlp undergoes independent, oscillating high-frequency phase variation in isogenic populations and can be expressed individually or concomitantly with other Vlps in a noncoordinate manner. All seven possible combinations of these three products were observed; however, no variants were found that lacked a Vlp. High-frequency size variation of each Vlp superimposed on combinatorial diversity in Vlp expression yields greater than 10(4) possible structurally distinct Vlp mosaics, of which 104 were documented along with 24 of 42 possible transitions among the seven Vlp combinations. In addition to these features, VlpA, VlpB, and VlpC were specifically recognized by serum antibodies from swine with experimental M. hyorhinis SK76-induced arthritis, indicating expression and immunogenicity of Vlps in the natural host. The structure and variation of Vlps and their known involvement in MAb-mediated modulation of mycoplasma-infected host cell properties and mycoplasma killing are discussed in relation to the surface architecture and adaptive potential of the wall-less mycoplasmas.

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Section: Discussionmentioning

confidence: 99%

The Vlp system of Mycoplasma hyorhinis: combinatorial expression of distinct size variant lipoproteins generating high-frequency surface antigenic variation

1991

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“…No sequence data of these proteins or their genes and no knowledge of the sequences involved in their expression are known. The only M. hyopneumoniae sequence data available up to now have been of rRNA genes which indicate that promoters of M. hyopneumoniae seem to be different from other mycoplasmas Taschke et al, , 1987, and two repeated sequences (Ferrell et al, 1989;Harasawa et al, 1991).…”

Section: Introductionmentioning

confidence: 99%

DNA sequence determination and biochemical analysis of the immunogenic protein P36, the lactate dehydrogenase (LDH) of Mycoplasma hyopneumoniae

Haldimann

Nicolet

Frey

1993

Journal of General Microbiology

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~The DNA sequence of the gene encoding the early and specific immunogenic protein P36 of Mycoplasmu hyopneumoniue has been determined. Comparison of the DNA sequence and the deduced amino acid sequence of P36 with known genes and proteins in data banks indicated that P36 is a L-lactate dehydrogenase (LDH) (EC 1.1.1.27). Biochemical analysis of protein P36 expressed from the cloned gene in Escherichiu coli confirmed that P36 has L-lactate dehydrogenase activity. Protein P36 of M. hyopneumoniae therefore is termed LDH and its gene Cdh. M. hyopneumoniae LDH was shown to contain the typical domains of LDH of other bacterial species. Immunologically however, we have shown that polyclonal antibodies against M. hyopneumoniue LDH do not crossreact with related LDH and show high specificity for M. hyopneunwniue. The ldh gene is preceded by several typical -10 sequences found in promoters of prokaryotes, but lacks the -35 sequence. Sequences rich in A + T, however, precede the -10 boxes, suggesting that factors involved in transcription initiation and their regulation may be different in M. hyopneumoniue compared to other bacterial species, but the putative ribosome binding site seems to be conserved.

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“…Double-stranded plasmid templates were isolated by the alkali lysis method (27) and purified by precipitation with 6.5% polyethylene glycol-0.8 M NaCl. Dideoxy sequencing reactions (28) (10). Lanes 1 to 6 and 8 to 11 represent chromosomal DNA from individual colony isolates of M. hyorhinis GDL-1, and lanes GDL and SK76 are from batch-passaged cultures of strains GDL-1 and SK76, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Diversification among the mycoplasmas is apparent in various morphological, biochemical, and serological characteristics of the organisms (11), with individual strains within several mycoplasma species displaying significant genotypic, phenotypic, and antigenic variability. In Mycoplasma hyorhinis, recently isolated gene sequences (10,29,38) have identified important genetic structures expected to be significant factors in generating such diversity. For example, repetition of mobile genetic elements, such as insertion sequences and transposons, distributed throughout the genome, provides a fertile source of DNA homologies for recombinational genomic reordering (21).…”

mentioning

confidence: 99%

“…For example, repetition of mobile genetic elements, such as insertion sequences and transposons, distributed throughout the genome, provides a fertile source of DNA homologies for recombinational genomic reordering (21). Although mycoplasmas have characteristically small genomes (24), several species, including M. hyorhinis (29), M. hyopneumoniae (10), M. fermentans (13), and M. pulmonis (3), contain multiple genomic copies of an insertion sequence related to IS3. It has not been established that interactions among such repetitive sequences are directly related to the high-frequency phenotypic variability within these species.…”

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An amplifiable DNA region from the Mycoplasma hyorhinis genome

Deng

McIntosh

1994

J Bacteriol

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A novel amplifiable genomic region that displays variability in the number of tandem copies of a 1,368-bp DNA sequence (designated RS-2) was discovered among individual clonal derivatives within Mycoplasma hyorhinis broth-grown cell populations. Clonal isolates representing variant subpopulations from the original broth culture were of a single size variant, and although continued culture under a variety of growth conditions did not result in further amplification of RS-2, evidence for deletion events which reduced RS-2 copy number, presumably by homologous recombination, was obtained. RS-2 homologous sequences were identified in all M. hyorhinis strains tested, but only the tissue culture-derived strains GDL-1 and GDL-2 showed variability in genomic dosage. The RS-2 nucleotide sequence established that each tandem copy is flanked by direct repeats of a 20-bp sequence and suggested a possible mechanism for its original duplication as the initial phase of a genetic amplification process. The coding strand was defined by PCR amplification of a reverse transcriptasegenerated cDNA, and its sequence revealed that RS-2 encodes a 456-residue internal, highly cysteine-rich domain of a larger M. hyorhinis protein whose coding sequence initiates and terminates in unique genomic sequences several hundred base pairs from RS-2 on either side of it. Changes in RS-2 copy number maintain the reading frame, and therefore the coding capacity, for this predicted size-variant protein.Mycoplasmas are a diverse group of nearly 100 bacterial species that were suggested to have evolved monophyletically at a high rate (17, 35, 37) from the low-G+C-content grampositive eubacterial phylogenetic branch (25). Their rapid evolutionary tempo reflects a high basal-level mutation rate (35, 37) and in several species may include a considerable frequency of more substantial genomic variabilities and rearrangements. Diversification among the mycoplasmas is apparent in various morphological, biochemical, and serological characteristics of the organisms (11), with individual strains within several mycoplasma species displaying significant genotypic, phenotypic, and antigenic variability. In Mycoplasma hyorhinis, recently isolated gene sequences (10, 29, 38) have identified important genetic structures expected to be significant factors in generating such diversity. For example, repetition of mobile genetic elements, such as insertion sequences and transposons, distributed throughout the genome, provides a fertile source of DNA homologies for recombinational genomic reordering (21). Although mycoplasmas have characteristically small genomes (24), several species, including M. hyorhinis (29), M. hyopneumoniae (10), M. fermentans (13), and M. pulmonis (3), contain multiple genomic copies of an insertion sequence related to IS3. It has not been established that interactions among such repetitive sequences are directly related to the high-frequency phenotypic variability within these species. However, their high copy number is likely to result in measura...

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A Mycoplasma genetic element resembling prokaryotic insertion sequences

Cited by 36 publications

References 49 publications

The Vlp system of Mycoplasma hyorhinis: combinatorial expression of distinct size variant lipoproteins generating high-frequency surface antigenic variation

The Vlp system of Mycoplasma hyorhinis: combinatorial expression of distinct size variant lipoproteins generating high-frequency surface antigenic variation

DNA sequence determination and biochemical analysis of the immunogenic protein P36, the lactate dehydrogenase (LDH) of Mycoplasma hyopneumoniae

An amplifiable DNA region from the Mycoplasma hyorhinis genome

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