Prevalence and Sequence Diversity of a Factor Required for Actin-Based Motility in Natural Populations of
            <i>Burkholderia</i>
            Species

Sitthidet, Chayada; Stevens, Joanne M.; Chantratita, Narisara; Currie, Bart J.; Peacock, Sharon J.; Korbsrisate, Sunee; Stevens, Mark P.

doi:10.1128/jcm.00368-08

Cited by 40 publications

(58 citation statements)

References 32 publications

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“…Intraspecies conservation of BimA was found to be high in natural populations of B. pseudomallei, B. mallei, and B. thailandensis (19,20), with the exception that a B. mallei-like BimA variant was found in B. pseudomallei isolates restricted to the Northern Territory of Australia, but not specific multilocus sequence types (19). ClustalW alignment of predicted BimA protein sequences indicated that tandem WH2 domains predicted to mediate the binding of actin monomers were conserved in sequenced B. pseudomallei strains (19). A proline-rich motif (IP 7 ; also defined as a PRM1 motif [reviewed in reference 9]) was also conserved.…”

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confidence: 99%

“…To identify conserved and variable portions of bimA, we previously surveyed the prevalence and sequence diversity of the gene in clinical and environmental isolates from areas of endemicity (19). Intraspecies conservation of BimA was found to be high in natural populations of B. pseudomallei, B. mallei, and B. thailandensis (19,20), with the exception that a B. mallei-like BimA variant was found in B. pseudomallei isolates restricted to the Northern Territory of Australia, but not specific multilocus sequence types (19).…”

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confidence: 99%

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Identification of Motifs of Burkholderia pseudomallei BimA Required for Intracellular Motility, Actin Binding, and Actin Polymerization

Sitthidet¹,

Korbsrisate²,

Layton³

et al. 2011

Journal of Bacteriology

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Actin-based motility of the melioidosis pathogen Burkholderia pseudomallei requires BimA (Burkholderia intracellular motility A). The mechanism by which BimA mediates actin assembly at the bacterial pole is ill-defined. Toward an understanding of the regions of B. pseudomallei BimA required for intracellular motility and the binding and polymerization of actin, we constructed plasmid-borne bimA variants and glutathione-Stransferase fusion proteins with in-frame deletions of specific motifs. A 13-amino-acid direct repeat and IP 7 proline-rich motif were dispensable for actin binding and assembly in vitro, and expression of the mutated proteins in a B. pseudomallei bimA mutant restored actin-based motility in J774.2 murine macrophage-like cells. However, two WASP homology 2 (WH2) domains were found to be required for actin binding, actin assembly, and plaque formation. A tract of five PDASX direct repeats influenced the polymerization of pyrene-actin monomers in vitro and was required for actin-based motility and intercellular spread, but not actin binding. None of the mutations impaired surface expression or polar targeting of BimA. The number of PDASX repeats varied in natural isolates from two to seven. Such repeats acted additively to promote pyrene-actin polymerization in vitro, with stepwise increases in the rate of polymerization as the number of repeats was increased. No differences in the efficiency of actin tail formation could be discerned between strains expressing BimA variants with two, five, or seven PDASX repeats. The data provide valuable new insights into the role of conserved and variable motifs of BimA in actin-based motility and intercellular spread of B. pseudomallei.

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Identification of Motifs of Burkholderia pseudomallei BimA Required for Intracellular Motility, Actin Binding, and Actin Polymerization

Sitthidet¹,

Korbsrisate²,

Layton³

et al. 2011

Journal of Bacteriology

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“…BimA is involved in actin polymerization, which is crucial for intracellular motility and intercellular spread and is thus a potential virulence factor. 12 Both patients with the B. mallei allele of bimA presented with septicemic pneumonia. While the B. mallei allele occurs in some Australian B. pseudomallei strains, Sitthidet and others 12 found none of 64 strains from across Southeast Asia harboring this allele that is present in B. mallei , the causative bacterium of glanders.…”

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confidence: 99%

“…12 Both patients with the B. mallei allele of bimA presented with septicemic pneumonia. While the B. mallei allele occurs in some Australian B. pseudomallei strains, Sitthidet and others 12 found none of 64 strains from across Southeast Asia harboring this allele that is present in B. mallei , the causative bacterium of glanders. To our knowledge the two Indian B. pseudomallei strains noted here to have the B. mallei -type bimA allele are the first B. pseudomallei isolates of Asian origin reported to contain this allele.…”

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confidence: 99%

Molecular Characterization of Clinical Burkholderia pseudomallei Isolates from India

Mukhopadhyay¹,

Kaestli²,

Vandana³

et al. 2011

The American Society of Tropical Medicine and Hygiene

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“…Another potential problem with the BimA assays is that B. pseudomallei strains from Australia also contain the N-terminus region of BimA, indicating that the BimA tests may not be suitable for specific B. mallei differentiation from Australian B. pseudomallei isolates (Sitthidet et al, 2008). In addition, B. thailandensis-like strain MSMB43 was detected by the BimA PCR assay .…”

Section: Flagellar Biosynthesis Proteinmentioning

confidence: 99%

PCR-based Methodologies Used to Detect and Differentiate theBurkholderia pseudomalleicomplex:B. pseudomallei,B. mallei, andB. thailandensis

Lowe

March²,

Bunnell³

et al. 2014

Current Issues in Molecular Biology

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Methods for the rapid detection and differentiation of the Burkholderia pseudomallei complex comprising B. pseudomallei, B. mallei, and B. thailandensis, have been the topic of recent research due to the high degree of phenotypic and genotypic similarities of these species. B. pseudomallei and B. mallei are recognized by the CDC as tier 1 select agents. The high mortality rates of glanders and melioidosis, their potential use as bioweapons, and their low infectious dose, necessitate the need for rapid and accurate detection methods. Although B. thailandensis is generally avirulent in mammals, this species displays very similar phenotypic characteristics to that of B. pseudomallei. Optimal identification of these species remains problematic, due to the difficulty in developing a sensitive, selective, and accurate assay. The development of PCR technologies has revolutionized diagnostic testing and these detection methods have become popular due to their speed, sensitivity, and accuracy. The purpose of this review is to provide a comprehensive overview and evaluation of the advancements in PCR-based detection and differentiation methodologies for the B. pseudomallei complex, and examine their potential uses in diagnostic and environmental testing.

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Prevalence and Sequence Diversity of a Factor Required for Actin-Based Motility in Natural Populations of Burkholderia Species

Cited by 40 publications

References 32 publications

Identification of Motifs of Burkholderia pseudomallei BimA Required for Intracellular Motility, Actin Binding, and Actin Polymerization

Identification of Motifs of Burkholderia pseudomallei BimA Required for Intracellular Motility, Actin Binding, and Actin Polymerization

Molecular Characterization of Clinical Burkholderia pseudomallei Isolates from India

PCR-based Methodologies Used to Detect and Differentiate theBurkholderia pseudomalleicomplex:B. pseudomallei,B. mallei, andB. thailandensis

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