Localized frameshift mutation generates selective, high-frequency phase variation of a surface lipoprotein encoded by a mycoplasma ABC transporter operon

Theiss, Patty; Wise, Kim S.

doi:10.1128/jb.179.12.4013-4022.1997

Cited by 59 publications

(52 citation statements)

References 64 publications

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“…However, because our studies were conducted with purified Phn ϩ isolates, we conclude that the phnE gene in E. coli K-12 behaves like a reversible gene switch, causing phase variation in phosphonate metabolism. This appears to be the first example of phase variation in a component of an ABC transporter in a gramnegative bacterium (10) although it is interesting that a highfrequency frameshift phase variation event affects the proposed substrate-binding lipoprotein encoded within an ABC transporter operon in Mycoplasma fermentans (20). It is not clear why "off" forms start to accumulate rapidly late in the growth cycle on phosphonates or once the selection for growth on phosphonates is removed.…”

Section: Discussionmentioning

confidence: 99%

Reversible Phase Variation in the phnE Gene, Which Is Required for Phosphonate Metabolism in Escherichia coli K-12

et al. 2004

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It is known that Escherichia coli K-12 is cryptic (Phn ؊ ) for utilization of methyl phosphonate (MePn) and that Phn ؉ variants can be selected for growth on MePn as the sole P source. Variants arise from deletion via a possible slip strand mechanism of one of three direct 8-bp repeat sequences in phnE, which restores function to a component of a putative ABC type transporter. Here we show that Phn ؉ variants are present at the surprisingly high frequency of >10 ؊2 in K-12 strains. Amplified-fragment length polymorphism analysis was used to monitor instability in phnE in various strains growing under different conditions. This revealed that, once selection for growth on MePn is removed, Phn ؉ revertants reappear and accumulate at high levels through reinsertion of the 8-bp repeat element sequence. It appears that, in K-12, phnE contains a highfrequency reversible gene switch, producing phase variation which either allows ("on" form) or blocks ("off" form) MePn utilization. The switch can also block usage of other metabolizable alkyl phosphonates, including the naturally occurring 2-aminoethylphosphonate. All K-12 strains, obtained from collections, appear in the "off" form even when bearing mutations in mutS, mutD, or dnaQ which are known to enhance slip strand events between repetitive sequences. The ability to inactivate the phnE gene appears to be unique to K-12 strains since the B strain is naturally Phn ؉ and lacks the inactivating 8-bp insertion in phnE, as do important pathogenic strains for which genome sequences are known and also strains isolated recently from environmental sources.Escherichia coli can use P III compounds such as phosphite and organophosphonates, e.g., methylphosphonate (MePn) and aminoethylphosphonate (AEPn) as P sources. Their metabolism involves the enzyme C-P lyase, which appears to have a relatively broad substrate specificity (21). Whereas the mechanism of C-P lyase is not well understood, much is known about a cluster of 17 contiguous phn genes in E. coli, required for utilization of P III compounds (5, 21). The phnGHIJK genes within this cluster are thought to encode the core components of C-P lyase, while phnF and phnO potentially encode regulatory proteins (5). Several phn genes appear to encode components of solute transporters, and it has been deduced that, among these, the phnCDE genes encode an ABC type transporter. In this transporter, phnC encodes the ABC permease component, phnD encodes the periplasmic binding protein, and phnE encodes the integral membrane component.An interesting feature of the genetics of phosphonate metabolism in E. coli is that the B strain can use phosphonates whereas the K-12 strain is cryptic despite containing the entire phn gene cluster (21). The genetic basis for this crypticity was investigated by Makino et al. (14) and traced to an 8-bp insertion in the coding region of the phnE gene in the K-12 strain relative to the B strain, causing truncation of the phnE product. They also observed that the 8-bp sequence is one element in the direct t...

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Section: Discussionmentioning

confidence: 99%

Reversible Phase Variation in the phnE Gene, Which Is Required for Phosphonate Metabolism in Escherichia coli K-12

et al. 2004

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“…This gene was identified as ptsG, which encodes a putative glucose PTS permease. Phase variation in mycoplasma transporters has so far only been documented for one subunit in an ABC transporter of Mycoplasma fermentans (Theiss & Wise, 1997). These authors speculated that variation might not only have consequences for immune evasion, but also provide alternative transport capacities through the use of different subsets of the same genes.…”

Section: Discussion a New Antigenic Variation In Mmymy Scmentioning

confidence: 99%

“…Protein variation not only enables mycoplasmas to escape the host immune defence system Neyrolles et al, 1999;Le Grand et al, 1996), but is also involved in adhesion (Sachse et al, 2000;Washburn et al, 1993), haemadsorption (Markham et al, 1993;Noormohammadi et al, 1997Noormohammadi et al, , 2000, membrane transport (Theiss & Wise, 1997) and immunomodulation (Muhlradt et al, 1998). The simplest form of surface variation is a reversible ON/OFF switch in protein expression, called phase variation.…”

Section: Introductionmentioning

confidence: 99%

“…The simplest form of surface variation is a reversible ON/OFF switch in protein expression, called phase variation. The wide range of mutations involved in such variation include promoter mutations (Yogev et al, 1991; Persson et al, 2002) or inversion (Horino et al, 2003), changes in putative transcription activators (Glew et al, 1998; Washburn et al, 1998), mutations that lead to gene truncation (Theiss & Wise, 1997), frameshift mutations (Boguslavsky et al, 2000; Winner et al, 2003;Zhang & Wise, 1997) and DNA inversions or gene conversions that fuse the coding gene to an active promoter (Bhugra et al, 1995;Glew et al, 2002;Noormohammadi et al, 2000;Lysnyansky et al, 2001b).Some surface proteins undergo antigenic variation by altering the size and/or epitope composition. Most of these proteins contain repetitive units of different structure.…”

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Variability of a glucose phosphotransferase system permease in Mycoplasma mycoides subsp. mycoides Small Colony

Gaurivaud¹,

Persson

Grand

et al. 2004

Microbiology

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Intraclonal antigenic variation in pathogenic mycoplasma species is considered an important feature of host-pathogen interaction. Such intraclonal protein variation was observed for the interaction of Mycoplasma mycoides subsp. mycoides Small Colony, the agent of contagious bovine pleuropneumonia, with mAb 3F3. Colony immunostaining allows the definition of 3F3 ON-and 3F3 OFF-type variants, which revert at low frequency. Targets of mAb 3F3 were shown to be surface located, and resided on multiple polypeptides in the 58-68 kDa size range. Phage display and a genomic database were combined to determine the gene encoding the proteins recognized by mAb 3F3. A gene encoding the putative permease of the glucose phosphotransferase system was identified. Genome sequence analysis of strain PG1 revealed two highly similar copies of this gene, resulting from duplication of the chromosomal region carrying the gene. Southern blot analysis demonstrated the presence of this duplication in almost every African strain tested, but not in European strains. DNA analysis revealed that ON/OFF switching is governed by a base substitution occurring upstream of the coding region for the 3F3 epitope. This event generates a stop codon that results in the premature termination of the PtsG protein. INTRODUCTIONMycoplasma mycoides subsp. mycoides biotype Small Colony (Mmymy SC) belongs to the class Mollicutes, trivial name mycoplasma, whose members are distinguished from other bacteria by their minute size and total lack of a rigid cell wall. Mmymy SC is the aetiological agent of contagious bovine pleuropneumonia (CBPP), a highly contagious respiratory disease in cattle. CBPP is the only bacterial disease included in the A list of the World Organization for Animal Health (OIE), which contains prioritized communicable animal diseases. During the nineteenth century, CBPP spread throughout the world, causing extensive losses in livestock. By prohibiting cattle movement, and by a programme of slaughtering and vaccination, CBPP was eradicated during the twentieth century, except in Africa and limited areas of Europe and Asia. Since the late 1980s, CBPP has become the major infectious disease affecting livestock in Africa, and the number of countries with CBPP-infected animals rose dramatically from 15 in the late 1970s to 26 in 2002 [Nicholas et al., 2000; OIE official data (www.oie.int)]. CBPP was thought to be almost eradicated from Europe by the early twentieth century, and until the 1960s only a few sporadic outbreaks continued to occur in the Iberian Peninsula. However, from 1980 onwards, hundreds of outbreaks were reported in France, Portugal, Spain and Italy. Control programmes were set up and no case of CBPP has been officially documented in Europe since 1999 (Portugal).Abbreviations: CBPP, contagious bovine pleuropneumonia; glucose PTS, phosphoenolpyruvate : glucose phosphotransferase system; HSA, human serum albumin; Mmymy SC, Mycoplasma mycoides subsp. mycoides biotype Small Colony. 0002-7247 G 2004 SGM Printed in Great Britain 4009Mic...

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“…This lipoprotein appears to be one of the haemagglutinins of this organism. Similar lipoproteins are present in other mycoplasmas, such as M. hyorrhinis (Rosengarten and Wise, 1990), M. hominis (Olson et al, 1991), M. arthritidis (Washburn et al, 1998), and M. fermentans (Theiss and Wise, 1997). M. fermentans through its M161Ag surface lipoprotein serves as a potent cytokine inducer for monocytes/macrophages and maturing dendritic cells, and it activates host complement (Muhlradt, 1997).…”

mentioning

confidence: 96%

Binding of mycoplasmas to solid phase adsorbents

Szathmáry¹,

Rajapakse²,

Székely

et al. 2005

Acta Veterinaria Hungarica

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The capture of mycoplasmas (M. hominis, M. buccale, M. fermentans, M. bovis, M. synoviae, M. gallisepticum and M. arthritidis) based on lipid structures and adhesion molecules present in the mycoplasmal membrane was tested using different chromatographic resins (ActiClean Etox, ClarEtox, Heparin-Actigel, Sulfated Hiflow and SulfEtox). All of the resins efficiently reduced mycoplasma concentrations in Phosphate Buffered Saline (PBS) and in Fetal Bovine Serum (FBS) by 3-8 logs in a few minutes. This technology could be used for removing mycoplasmas from tissue culture components such as serum, and for concentrating mycoplasmas in vaccine production.

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Localized frameshift mutation generates selective, high-frequency phase variation of a surface lipoprotein encoded by a mycoplasma ABC transporter operon

Cited by 59 publications

References 64 publications

Reversible Phase Variation in the phnE Gene, Which Is Required for Phosphonate Metabolism in Escherichia coli K-12

Reversible Phase Variation in the phnE Gene, Which Is Required for Phosphonate Metabolism in Escherichia coli K-12

Variability of a glucose phosphotransferase system permease in Mycoplasma mycoides subsp. mycoides Small Colony

Binding of mycoplasmas to solid phase adsorbents

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