The effect of Wag31 phosphorylation on the cells and the cell envelope fraction of wild-type and conditional mutants of Mycobacterium smegmatis studied by visible-wavelength Raman spectroscopy

Hamasha, Khozima; Sahana, M.; Jani, Charul; Nyayapathy, Seeta; Kang, Choong-Min; Rehse, Steven J.

doi:10.1016/j.bbrc.2009.11.117

Cited by 16 publications

(14 citation statements)

References 11 publications

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“…Consistent with this finding, we recently found in a Raman spectroscopic analyses that cells containing wag31T73E Mtb allele had increased intensity of the Raman peaks that have previously been attributed to D-glutamic acid, D-alanine, and N -acetylglucosamine components of peptidoglycan [23,24] than cells expressing wild-type wag31 Mtb , which in turn showed higher intensity of the these peaks than cells with wag31T73A Mtb [25]. An increase in the intensity of these peaks suggests an increase in the quantity of these molecules, thus peptidoglycan in the cell.…”

Section: Resultssupporting

confidence: 63%

Regulation of Polar Peptidoglycan Biosynthesis by Wag31 Phosphorylation in Mycobacteria

et al. 2010

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BackgroundSensing and responding to environmental changes is a central aspect of cell division regulation. Mycobacterium tuberculosis contains eleven Ser/Thr kinases, two of which, PknA and PknB, are key signaling molecules that regulate cell division/morphology. One substrate of these kinases is Wag31, and we previously showed that partial depletion of Wag31 caused morphological changes indicative of cell wall defects, and that the phosphorylation state of Wag31 affected cell growth in mycobacteria. In the present study, we further characterized the role of the Wag31 phosphorylation in polar peptidoglycan biosynthesis.ResultsWe demonstrate that the differential growth among cells expressing different wag31 alleles (wild-type, phosphoablative, or phosphomimetic) is caused by, at least in part, dissimilar nascent peptidoglycan biosynthesis. The phosphorylation state of Wag31 is found to be important for protein-protein interactions between the Wag31 molecules, and thus, for its polar localization. Consistent with these results, cells expressing a phosphomimetic wag31 allele have a higher enzymatic activity in the peptidoglycan biosynthetic pathway.ConclusionsThe Wag31Mtb phosphorylation is a novel molecular mechanism by which Wag31Mtb regulates peptidoglycan synthesis and thus, optimal growth in mycobacteria.

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Section: Resultssupporting

confidence: 63%

Regulation of Polar Peptidoglycan Biosynthesis by Wag31 Phosphorylation in Mycobacteria

et al. 2010

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“…The phosphorylation state of Wag31 is modulated by the serine/theronine kinases PknA and PknB [33, 35–37]. Overexpression of wild-type or a phosphomimetic allele of Wag31, but not a phosphoablative allele, leads to the production of short, rounded cells [37].…”

Section: Polar Growth Patterns In Specific Bacterial Taxamentioning

confidence: 99%

“…Expression of a phosphomimetic allele of Wag31 or overexpression of PknB, which increases the pool of phosphorylated Wag31, leads to an increase in polar localization of Wag31 and a subsequent increase in nascent peptidoglycan synthesis [36]. Furthermore, expression of the phosphomimetic allele significantly increases the combined activity of MraY and MurG, transferases involved in the final step of peptidoglycan synthesis [36] and leads to an elevated amount of D-glutamic acid, D-alanine, and N-acetylglucosamine components of peptidoglycan [35]. These results suggest the Wag31 phosphorylation regulates the activity of the peptidoglycan biosynthesis machinery, although it remains unknown if the regulation is direct or indirect.…”

Section: Polar Growth Patterns In Specific Bacterial Taxamentioning

confidence: 99%

Polarity and the diversity of growth mechanisms in bacteria

Brown

Kysela

Brun

2011

Seminars in Cell & Developmental Biology

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Bacterial cell growth is a complex process consisting of two distinct phases: cell elongation and septum formation prior to cell division. Although bacteria have evolved several different mechanisms for cell growth, it is clear that tight spatial and temporal regulation of peptidoglycan synthesis is a common theme. In this review, we discuss bacterial cell growth with a particular emphasis on bacteria that utilize tip extension as a mechanism for cell elongation. We describe polar growth among diverse bacteria and consider the advantages and consequences of this mode of cell elongation.

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“…A previous study of the chromosome sequence of Hfr K-12 and O157:H7 revealed that O157:H7 has evolved from Hfr K-12 after the attack of a lysogenic bacteriophage [19,20]. We have previously utilized RS to study the uptake of the 5-carbon polyol xylitol by several of these E. coli strains [21] as well as to measure the molecular differences of conditional mutants of Mycobacterium smegmatis expressing three different alleles relating to the phosphorylation of Wag31, a key cell-division protein [22]. …”

Section: Introductionmentioning

confidence: 99%

Sensitive and specific discrimination of pathogenic and nonpathogenic Escherichia coli using Raman spectroscopy—a comparison of two multivariate analysis techniques

Hamasha¹,

Mohaidat²,

Putnam³

et al. 2013

Biomed. Opt. Express

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The determination of bacterial identity at the strain level is still a complex and time-consuming endeavor. In this study, visible wavelength spontaneous Raman spectroscopy has been used for the discrimination of four closely related Escherichia coli strains: pathogenic enterohemorrhagic E. coli O157:H7 and non-pathogenic E. coli C, E. coli Hfr K-12, and E. coli HF4714. Raman spectra from 600 to 2000 cm−1 were analyzed with two multivariate chemometric techniques, principal component-discriminant function analysis and partial least squares-discriminant analysis, to determine optimal parameters for the discrimination of pathogenic E. coli from the non-pathogenic strains. Spectral preprocessing techniques such as smoothing with windows of various sizes and differentiation were investigated. The sensitivity and specificity of both techniques was in excess of 95%, determined by external testing of the chemometric models. This study suggests that spontaneous Raman spectroscopy with visible wavelength excitation is potentially useful for the rapid identification and classification of clinically-relevant bacteria at the strain level.

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The effect of Wag31 phosphorylation on the cells and the cell envelope fraction of wild-type and conditional mutants of Mycobacterium smegmatis studied by visible-wavelength Raman spectroscopy

Cited by 16 publications

References 11 publications

Regulation of Polar Peptidoglycan Biosynthesis by Wag31 Phosphorylation in Mycobacteria

Regulation of Polar Peptidoglycan Biosynthesis by Wag31 Phosphorylation in Mycobacteria

Polarity and the diversity of growth mechanisms in bacteria

Sensitive and specific discrimination of pathogenic and nonpathogenic Escherichia coli using Raman spectroscopy—a comparison of two multivariate analysis techniques

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