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Mycoplasma pneumoniae large DNA repetitive elements RepMP1 show type specific organization among strains.

Musatovova O, Kannan TR, Baseman JB - PLoS ONE (2012)

Bottom Line: Recently, we described RepMP1-associated sequence variations between reference strain M129 and clinical isolate S1 that involved three RepMP1-genes (i.e. mpn130, mpn137 and mpn138).The only other detected difference was varying numbers of the 21-nucleotide tandem repeats within mpn127, mpn137, mpn501 and mpn524.Furthermore, typing of strains through analysis of large RepMPs localized within the adhesin P1 operon revealed that sequence divergence involving RepMP1-genes mpn130, mpn137 and mpn138 is strictly type-specific.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, The University of Texas Health Science Center at San Antonio, San Antonio, Texas, United States of America. musatovova@gmail.com

ABSTRACT
Mycoplasma pneumoniae is the smallest self-replicating bacterium with a streamlined genome of 0.81 Mb. Complete genome analysis revealed the presence of multiple copies of four large repetitive elements (designated RepMP1, RepMP2/3, RepMP4 and RepMP5) that are implicated in creating sequence variations among individual strains. Recently, we described RepMP1-associated sequence variations between reference strain M129 and clinical isolate S1 that involved three RepMP1-genes (i.e. mpn130, mpn137 and mpn138). Using PCR and sequencing we analyze 28 additional M. pneumoniae strains and demonstrate the existence of S1-like sequence variants in nine strains and M129-like variants in the remaining nineteen strains. We propose a series of recombination steps that facilitates transition from M129- to S1-like sequence variants. Next we examined the remaining RepMP1-genes and observed no other rearrangements related to the repeat element. The only other detected difference was varying numbers of the 21-nucleotide tandem repeats within mpn127, mpn137, mpn501 and mpn524. Furthermore, typing of strains through analysis of large RepMPs localized within the adhesin P1 operon revealed that sequence divergence involving RepMP1-genes mpn130, mpn137 and mpn138 is strictly type-specific. Once more our analysis confirmed existence of two highly conserved groups of M. pneumoniae strains.

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Type-specific deletion of MPN130.A. PCR amplification of MPN129 to MPN131 regions. Chromosomal DNA of four type 1 and eight type 2 strains was used and generated products were visualized on 1% agarose for analysis. Amplicon size was estimated using 1-kb DNA ladder. B. Comparison of MPN129-MPN131 regions among M. pneumoniae clinical strains. In reference strain M129 and other type 1 isolates, the RepMP1-containing gene (MPN130, orange arrow) is located between coding regions MPN129 and MPN131. In all tested type 2 strains, MPN130 is missing. The location of RepMP1-core element and sRepB within analyzed regions is indicated. The position of amplified genomic regions (line) and primers (▸,◂) is shown for both type 1 and type 2 strains, and the region deleted in type 2 strains is presented (striped bar).
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pone-0047625-g002: Type-specific deletion of MPN130.A. PCR amplification of MPN129 to MPN131 regions. Chromosomal DNA of four type 1 and eight type 2 strains was used and generated products were visualized on 1% agarose for analysis. Amplicon size was estimated using 1-kb DNA ladder. B. Comparison of MPN129-MPN131 regions among M. pneumoniae clinical strains. In reference strain M129 and other type 1 isolates, the RepMP1-containing gene (MPN130, orange arrow) is located between coding regions MPN129 and MPN131. In all tested type 2 strains, MPN130 is missing. The location of RepMP1-core element and sRepB within analyzed regions is indicated. The position of amplified genomic regions (line) and primers (▸,◂) is shown for both type 1 and type 2 strains, and the region deleted in type 2 strains is presented (striped bar).

Mentions: We reported that the 49-nucleotide region identified within mpn138/7 matched completely a region within mpn130, a gene that was absent in clinical strain S1 [5]. Chromosomal regions containing mpn130 and the adjacent genes (Figure 2) were therefore amplified from all strains and compared to M129 (2289 bp) and S1 (1609 bp) amplicons. All M129-like strains yielded ∼2.3 kb size product and all S1-like strains generated ∼1.6 kb size product (Figure 2A). Subsequent sequencing of the 1.6 kb amplicons revealed the identical 680-bp deletion in all tested S1-like strains (enclosing mpn130 together with 116 nucleotides of up- and 141 nucleotides of downstream regions; Figure 2B) [5].


Mycoplasma pneumoniae large DNA repetitive elements RepMP1 show type specific organization among strains.

Musatovova O, Kannan TR, Baseman JB - PLoS ONE (2012)

Type-specific deletion of MPN130.A. PCR amplification of MPN129 to MPN131 regions. Chromosomal DNA of four type 1 and eight type 2 strains was used and generated products were visualized on 1% agarose for analysis. Amplicon size was estimated using 1-kb DNA ladder. B. Comparison of MPN129-MPN131 regions among M. pneumoniae clinical strains. In reference strain M129 and other type 1 isolates, the RepMP1-containing gene (MPN130, orange arrow) is located between coding regions MPN129 and MPN131. In all tested type 2 strains, MPN130 is missing. The location of RepMP1-core element and sRepB within analyzed regions is indicated. The position of amplified genomic regions (line) and primers (▸,◂) is shown for both type 1 and type 2 strains, and the region deleted in type 2 strains is presented (striped bar).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3472980&req=5

pone-0047625-g002: Type-specific deletion of MPN130.A. PCR amplification of MPN129 to MPN131 regions. Chromosomal DNA of four type 1 and eight type 2 strains was used and generated products were visualized on 1% agarose for analysis. Amplicon size was estimated using 1-kb DNA ladder. B. Comparison of MPN129-MPN131 regions among M. pneumoniae clinical strains. In reference strain M129 and other type 1 isolates, the RepMP1-containing gene (MPN130, orange arrow) is located between coding regions MPN129 and MPN131. In all tested type 2 strains, MPN130 is missing. The location of RepMP1-core element and sRepB within analyzed regions is indicated. The position of amplified genomic regions (line) and primers (▸,◂) is shown for both type 1 and type 2 strains, and the region deleted in type 2 strains is presented (striped bar).
Mentions: We reported that the 49-nucleotide region identified within mpn138/7 matched completely a region within mpn130, a gene that was absent in clinical strain S1 [5]. Chromosomal regions containing mpn130 and the adjacent genes (Figure 2) were therefore amplified from all strains and compared to M129 (2289 bp) and S1 (1609 bp) amplicons. All M129-like strains yielded ∼2.3 kb size product and all S1-like strains generated ∼1.6 kb size product (Figure 2A). Subsequent sequencing of the 1.6 kb amplicons revealed the identical 680-bp deletion in all tested S1-like strains (enclosing mpn130 together with 116 nucleotides of up- and 141 nucleotides of downstream regions; Figure 2B) [5].

Bottom Line: Recently, we described RepMP1-associated sequence variations between reference strain M129 and clinical isolate S1 that involved three RepMP1-genes (i.e. mpn130, mpn137 and mpn138).The only other detected difference was varying numbers of the 21-nucleotide tandem repeats within mpn127, mpn137, mpn501 and mpn524.Furthermore, typing of strains through analysis of large RepMPs localized within the adhesin P1 operon revealed that sequence divergence involving RepMP1-genes mpn130, mpn137 and mpn138 is strictly type-specific.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, The University of Texas Health Science Center at San Antonio, San Antonio, Texas, United States of America. musatovova@gmail.com

ABSTRACT
Mycoplasma pneumoniae is the smallest self-replicating bacterium with a streamlined genome of 0.81 Mb. Complete genome analysis revealed the presence of multiple copies of four large repetitive elements (designated RepMP1, RepMP2/3, RepMP4 and RepMP5) that are implicated in creating sequence variations among individual strains. Recently, we described RepMP1-associated sequence variations between reference strain M129 and clinical isolate S1 that involved three RepMP1-genes (i.e. mpn130, mpn137 and mpn138). Using PCR and sequencing we analyze 28 additional M. pneumoniae strains and demonstrate the existence of S1-like sequence variants in nine strains and M129-like variants in the remaining nineteen strains. We propose a series of recombination steps that facilitates transition from M129- to S1-like sequence variants. Next we examined the remaining RepMP1-genes and observed no other rearrangements related to the repeat element. The only other detected difference was varying numbers of the 21-nucleotide tandem repeats within mpn127, mpn137, mpn501 and mpn524. Furthermore, typing of strains through analysis of large RepMPs localized within the adhesin P1 operon revealed that sequence divergence involving RepMP1-genes mpn130, mpn137 and mpn138 is strictly type-specific. Once more our analysis confirmed existence of two highly conserved groups of M. pneumoniae strains.

Show MeSH
Related in: MedlinePlus