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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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Sequence variations involving MPN137 and MPN138.A. PCR amplification of MPN138/7 regions. Chromosomal DNA of four type 1 and eight type 2 strains was used, and generated products were electrophoresed on 1% agarose. Amplicon size was estimated using 1-kb DNA ladder. B. Comparison of MPN137 and MPN138 regions in different clinical strains. The analyzed region contains loci MPN139 (open arrow), MPN138 (red arrow) and MPN137 (blue arrow) in M129 strain and all tested type 1 isolates. The fused reading frame (MPN138/7, red and blue arrow) was found in all tested type 2 strains. The location of RepMP1-core elements and short repeats B and C within analyzed regions is indicated. The position of amplified genomic regions (line) and primers (▸,◂) is shown for both type 1 and type 2 strains. The region deleted in type 2 strains is presented (striped bar) and the 49 nt-region originally not found in either MPN137 or MPN138 is indicated (orange stripe, **).
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pone-0047625-g001: Sequence variations involving MPN137 and MPN138.A. PCR amplification of MPN138/7 regions. Chromosomal DNA of four type 1 and eight type 2 strains was used, and generated products were electrophoresed on 1% agarose. Amplicon size was estimated using 1-kb DNA ladder. B. Comparison of MPN137 and MPN138 regions in different clinical strains. The analyzed region contains loci MPN139 (open arrow), MPN138 (red arrow) and MPN137 (blue arrow) in M129 strain and all tested type 1 isolates. The fused reading frame (MPN138/7, red and blue arrow) was found in all tested type 2 strains. The location of RepMP1-core elements and short repeats B and C within analyzed regions is indicated. The position of amplified genomic regions (line) and primers (▸,◂) is shown for both type 1 and type 2 strains. The region deleted in type 2 strains is presented (striped bar) and the 49 nt-region originally not found in either MPN137 or MPN138 is indicated (orange stripe, **).

Mentions: We set out to categorize sequence variations among a set of 30 M. pneumoniae strains by amplifying chromosomal regions containing mpn138 and mpn137 genes. The assembled set included reference strains B9-M129 (type 1), FH (type 2) [9], other M. pneumoniae strains deposited in ATCC as well as clinical strains of different geographic origins collected at different times by us and others (Table 2). PI1428, PN -and U-series generated a ∼2.6 kb PCR product similar to M129 whereas L2, SA1, FH, Mac, R32P, UTMB-10, and strains of TW series yielded ∼1.6 kb amplicons that matched with a PCR product generated for S1 isolate (Figure 1A).


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

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

Sequence variations involving MPN137 and MPN138.A. PCR amplification of MPN138/7 regions. Chromosomal DNA of four type 1 and eight type 2 strains was used, and generated products were electrophoresed on 1% agarose. Amplicon size was estimated using 1-kb DNA ladder. B. Comparison of MPN137 and MPN138 regions in different clinical strains. The analyzed region contains loci MPN139 (open arrow), MPN138 (red arrow) and MPN137 (blue arrow) in M129 strain and all tested type 1 isolates. The fused reading frame (MPN138/7, red and blue arrow) was found in all tested type 2 strains. The location of RepMP1-core elements and short repeats B and C within analyzed regions is indicated. The position of amplified genomic regions (line) and primers (▸,◂) is shown for both type 1 and type 2 strains. The region deleted in type 2 strains is presented (striped bar) and the 49 nt-region originally not found in either MPN137 or MPN138 is indicated (orange stripe, **).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3472980&req=5

pone-0047625-g001: Sequence variations involving MPN137 and MPN138.A. PCR amplification of MPN138/7 regions. Chromosomal DNA of four type 1 and eight type 2 strains was used, and generated products were electrophoresed on 1% agarose. Amplicon size was estimated using 1-kb DNA ladder. B. Comparison of MPN137 and MPN138 regions in different clinical strains. The analyzed region contains loci MPN139 (open arrow), MPN138 (red arrow) and MPN137 (blue arrow) in M129 strain and all tested type 1 isolates. The fused reading frame (MPN138/7, red and blue arrow) was found in all tested type 2 strains. The location of RepMP1-core elements and short repeats B and C within analyzed regions is indicated. The position of amplified genomic regions (line) and primers (▸,◂) is shown for both type 1 and type 2 strains. The region deleted in type 2 strains is presented (striped bar) and the 49 nt-region originally not found in either MPN137 or MPN138 is indicated (orange stripe, **).
Mentions: We set out to categorize sequence variations among a set of 30 M. pneumoniae strains by amplifying chromosomal regions containing mpn138 and mpn137 genes. The assembled set included reference strains B9-M129 (type 1), FH (type 2) [9], other M. pneumoniae strains deposited in ATCC as well as clinical strains of different geographic origins collected at different times by us and others (Table 2). PI1428, PN -and U-series generated a ∼2.6 kb PCR product similar to M129 whereas L2, SA1, FH, Mac, R32P, UTMB-10, and strains of TW series yielded ∼1.6 kb amplicons that matched with a PCR product generated for S1 isolate (Figure 1A).

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