Raman spectroscopic typing reveals the presence of carotenoids in Mycoplasma pneumoniae

Maquelin, Kees; Hoogenboezem, Theo; Jachtenberg, Jan-willem; Dumke, Roger; Jacobs, Enno; Puppels, Gerwin J.; Hartwig, Nico G.; Vink, Cornelis

doi:10.1099/mic.0.026724-0

Cited by 28 publications

(24 citation statements)

References 43 publications

(45 reference statements)

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“…All strains, except strain R003, have been described by Dorigo-Zetsma et al (2001), and were isolated in Denmark and The Netherlands between 1962 and 1995. Strain R003 was isolated in Germany in 1991 and has been described previously by Dumke et al (2003) and Maquelin et al (2009). The complete genome sequence of M. pneumoniae strain M129 (ATCC 29342; a subtype 1 strain) was used as a reference (Dandekar et al, 2000;Himmelreich et al, 1996).…”

Section: Methodsmentioning

confidence: 99%

Variation in a surface-exposed region of the Mycoplasma pneumoniae P40 protein as a consequence of homologous DNA recombination between RepMP5 elements

et al. 2011

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Mycoplasma pneumoniae is a human pathogen that causes a range of respiratory tract infections. The first step in infection is adherence of the bacteria to the respiratory epithelium. This step is mediated by a specialized organelle, which contains several proteins (cytadhesins) that have an important function in adherence. Two of these cytadhesins, P40 and P90, represent the proteolytic products from a single 130 kDa protein precursor, which is encoded by the MPN142 gene. Interestingly, MPN142 contains a repetitive DNA element, termed RepMP5, of which homologues are found at seven other loci within the M. pneumoniae genome. It has been hypothesized that these RepMP5 elements, which are similar but not identical in sequence, recombine with their counterpart within MPN142 and thereby provide a source of sequence variation for this gene. As this variation may give rise to amino acid changes within P40 and P90, the recombination between RepMP5 elements may constitute the basis of antigenic variation and, possibly, immune evasion by M. pneumoniae. To investigate the sequence variation of MPN142 in relation to inter-RepMP5 recombination, we determined the sequences of all RepMP5 elements in a collection of 25 strains. The results indicate that: (i) inter-RepMP5 recombination events have occurred in seven of the strains, and (ii) putative RepMP5 recombination events involving MPN142 have induced amino acid changes in a surface-exposed part of the P40 protein in two of the strains. We conclude that recombination between RepMP5 elements is a common phenomenon that may lead to sequence variation of MPN142-encoded proteins.

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Section: Methodsmentioning

confidence: 99%

Variation in a surface-exposed region of the Mycoplasma pneumoniae P40 protein as a consequence of homologous DNA recombination between RepMP5 elements

et al. 2011

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“…15531), and MAC (ATCC no. 15492), as well as clinical strains R035, P05/132, M688/98, and T79 (20), were cultured in Mycoplasma medium, as described previously (12).…”

Section: Methodsmentioning

confidence: 99%

Macrolide Resistance Determination and Molecular Typing of Mycoplasma pneumoniae in Respiratory Specimens Collected between 1997 and 2008 in The Netherlands

et al. 2012

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dAn important role in the treatment regimens for Mycoplasma pneumoniae infections is played by macrolide (ML) antibiotics. In the past few years, however, a steady increase has been detected in the worldwide prevalence of ML-resistant (ML r ) M. pneumoniae strains. It is obvious that this increase necessitates a continuous monitoring of ML r and, when detected, modification of antibiotic treatment modalities. Previously, we developed a pyrosequencing-based assay system for the genetic determination of ML r as well as molecular typing of M. pneumoniae. In this study, the sensitivity of this system was improved by the inclusion of a nested-PCR protocol. The modified system was applied to 114 M. pneumoniae-positive specimens that were obtained from a collection of 4,390 samples from patients with acute respiratory tract infections. These samples were collected between 1997 and 2008 in The Netherlands. The pyrosequencing system produced reliable data in 86% of the specimens that contained >500 M. pneumoniae genome copies/ml of patient sample. Each of these samples contained DNA of the ML-sensitive genotype. While 43% of the samples were found to harbor the M. pneumoniae subtype 1 genotype, 57% contained the subtype 2 genotype. We conclude that the pyrosequencing-based assay system is a useful tool for ML r determination and molecular typing of M. pneumoniae in patient samples. ML r -associated M. pneumoniae genotypes, however, were not found in the current study population. Mycoplasma pneumoniae is a pathogen of the human respiratory tract and one of the most prevalent causes of community-acquired pneumonia. General treatment options for M. pneumoniae infections include macrolides (MLs), tetracyclines, and fluoroquinolones. In young children, however, tetracyclines and fluoroquinolones are contraindicated due to unfavorable effects, such as discoloration of teeth by tetracyclines and putative cartilage damage induced by fluoroquinolones. The antibiotics of choice for this patient group are therefore ML antibiotics, such as clarithromycin and azithromycin. In the past decade, however, increasing numbers of M. pneumoniae clinical isolates were found to be resistant against these antibiotics. ML resistance (ML r ) was first reported in Japan in 2002 (14), and since then it has also been detected in the United States (10, 26), Europe (3,7,17,18), eastern Asia (13), and the Middle East (1). The prevalence of ML r varies widely, from 1 to 2% in Denmark (18) to as high as 90% in China (11). Clearly, the emergence of ML r poses a significant threat to the use of MLs in the treatment of M. pneumoniae infections. It is therefore important that ML r among M. pneumoniae isolates is rapidly and efficiently monitored in order to allow effective antibiotic treatment.Although the sensitivity of M. pneumoniae to antibiotics can be determined unambiguously on cultured bacterial isolates, the culturing of M. pneumoniae is time-consuming and precludes the timely identification of antibiotic-resistant isolates. The monitoring of ML r in...

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“…All M. pneumoniae strains used in this study, including reference strains FH (ATCC 15531), M129 (ATCC 29342), and PI 1428 (ATCC 29085), as well as M. genitalium strain G37 (ATCC 33530), were cultured in Mycoplasma medium, as described previously (18).…”

Section: Methodsmentioning

confidence: 99%

Functional Characterization of the RuvB Homologs from Mycoplasma pneumoniae and Mycoplasma genitalium

Estevão¹,

Sluijter²,

Hartwig³

et al. 2011

Journal of Bacteriology

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Homologous recombination between repeated DNA elements in the genomes of Mycoplasma species has been hypothesized to be a crucial causal factor in sequence variation of antigenic proteins at the bacterial surface. To investigate this notion, studies were initiated to identify and characterize the proteins that form part of the homologous DNA recombination machinery in Mycoplasma pneumoniae as well as Mycoplasma genitalium. A significant portion of the genomes of Mycoplasma pneumoniae and Mycoplasma genitalium consists of repeated DNA elements. This is remarkable as the genomes of these closely related human pathogens are relatively small, with lengths of 816 kb and 580 kb, respectively (6, 8). In M. pneumoniae, the repeated DNA elements are termed RepMP elements (29,37,44), whereas in M. genitalium, they are referred to as MgPa repeats (MgPars) (6,26,27). Although the RepMP elements and MgPa repeats do not have significant sequence homology, they do share two important features: (i) the different variants of these elements are similar in sequence but not identical, and (ii) one or more of these variants form part of open reading frames (ORFs) that code for antigenic surface proteins, such as the P1 protein in M. pneumoniae and the MgPa protein in M. genitalium. Because both P1 and MgPa can display sequence variation in the regions encoded by the RepMP or MgPar elements, it has been hypothesized that this variation is caused by homologous DNA recombination between the different variants of the RepMP or MgPar elements (10-11, 15, 17, 35, 36). Obviously, these recombination processes may supply a plethora of sequence variation to the P1 and MgPa genes, which can subsequently lead to amino acid sequence variation of the encoded antigenic proteins. Homologous DNA recombination between the repeated DNA elements in both Mycoplasma species may thus contribute significantly to evasion of these pathogens from the host's immune system.It is probable that the enzymatic machinery that governs recombination between repeated DNA elements in M. pneumoniae and M. genitalium largely overlaps with the machinery that is involved in homologous DNA recombination and DNA repair in these bacteria. Sequence analysis of the genomes of both M. pneumoniae and M. genitalium has revealed the presence of a distinct and limited set of ORFs that may encode the core proteins involved in DNA recombination pathways (2,6,8). The activities of some of these putative core proteins have already been investigated. These proteins include RuvA and a single-stranded DNA (ssDNA)-binding protein (SSB) from M. pneumoniae (9, 32) and the RecA and RecU proteins from both M. pneumoniae and M. genitalium (31,33,34). While the first three proteins exhibited activities that are similar to those of their counterparts from other bacteria, the RecU proteins from both Mycoplasma species displayed unusual and unique characteristics. First, while RecU from M. genitalium (RecU Mge ) was found to bind and cleave Holliday junction (HJ) substrates in a specific fashion, th...

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Raman spectroscopic typing reveals the presence of carotenoids in Mycoplasma pneumoniae

Cited by 28 publications

References 43 publications

Variation in a surface-exposed region of the Mycoplasma pneumoniae P40 protein as a consequence of homologous DNA recombination between RepMP5 elements

Variation in a surface-exposed region of the Mycoplasma pneumoniae P40 protein as a consequence of homologous DNA recombination between RepMP5 elements

Macrolide Resistance Determination and Molecular Typing of Mycoplasma pneumoniae in Respiratory Specimens Collected between 1997 and 2008 in The Netherlands

Functional Characterization of the RuvB Homologs from Mycoplasma pneumoniae and Mycoplasma genitalium

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