Identification of procaryotic developmental stages by statistical analyses of two‐dimensional gel patterns

Vohradský, Jiřı́; Li, Xinming; Thompson, Charles J.

doi:10.1002/elps.1150180817

Cited by 51 publications

(41 citation statements)

References 16 publications

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“…2D gel electrophoresis was performed as described by Vohradsky et al (1997). The IEF of proteins in the first dimension was carried out using 24 cm Immobiline DryStrips, pH 4-7 (Amersham Pharmacia) followed by electrophoretic separation on 10 % SDS-PAGE gels in the second dimension.…”

Section: Methodsmentioning

confidence: 99%

Morphological changes and proteome response of Corynebacterium glutamicum to a partial depletion of FtsI

et al. 2006

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In Corynebacterium glutamicum, as in many Gram-positive bacteria, the cell division gene ftsI is located at the beginning of the dcw cluster, which comprises cell division-and cell wall-related genes. Transcriptional analysis of the cluster revealed that ftsI is transcribed as part of a polycistronic mRNA, which includes at least mraZ, mraW, ftsL, ftsI and murE, from a promoter that is located upstream of mraZ. ftsI appears also to be expressed from a minor promoter that is located in the intergenic ftsL-ftsI region. It is an essential gene in C. glutamicum, and a reduced expression of ftsI leads to the formation of larger and filamentous cells. A translational GFP-FtsI fusion protein was found to be functional and localized to the mid-cell of a growing bacterium, providing evidence of its role in cell division in C. glutamicum. This study involving proteomic analysis (using 2D SDS-PAGE) of a C. glutamicum strain that has partially depleted levels of FtsI reveals that at least 20 different proteins were overexpressed in the organism. Eight of these overexpressed proteins, which include DivIVA, were identified by MALDI-TOF. Overexpression of DivIVA was confirmed by Western blotting using anti-DivIVA antibodies, and also by fluorescence microscopy analysis of a C. glutamicum RESF1 strain expressing a chromosomal copy of a divIVA-gfp transcriptional fusion. Overexpression of DivIVA was not observed when FtsI was inhibited by cephalexin treatment or by partial depletion of FtsZ.

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Section: Methodsmentioning

confidence: 99%

Morphological changes and proteome response of Corynebacterium glutamicum to a partial depletion of FtsI

et al. 2006

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“…The sensitivity range of the film was expanded by merging two different exposures of a single gel. The films were scanned using a Molecular Dynamics laser scanner and processed with pdQuest gel analysis software (30). Under these conditions, the electropherogram was able to reliably resolve more than 1,000 protein spots.…”

Section: Methodsmentioning

confidence: 99%

“…The developmental database recorded changes in a culture radiolabeled at 16 different time points (12,16,20,24,26,28,30,32,36,40,44,48,54,60,66, and 72 h; Fig. 1) (14).…”

Section: Methodsmentioning

confidence: 99%

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Developmental Control of Stress Stimulons in Streptomyces coelicolor Revealed by Statistical Analyses of Global Gene Expression Patterns

Vohradský

Dale

et al. 2000

J Bacteriol

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Stress-induced regulatory networks coordinated with a procaryotic developmental program were revealed by two-dimensional gel analyses of global gene expression. Four developmental stages were identified by their distinctive protein synthesis patterns using principal component analysis. Statistical analyses focused on five stress stimulons (induced by heat, cold, salt, ethanol, or antibiotic shock) and their synthesis during development. Unlike other bacteria, for which various stresses induce expression of similar sets of protein spots, in Streptomyces coelicolor heat, salt, and ethanol stimulons were composed of independent sets of proteins. This suggested independent control by different physiological stress signals and their corresponding regulatory systems. These stress proteins were also under developmental control. Cluster analysis of stress protein synthesis profiles identified 10 different developmental patterns or "synexpression groups." Proteins induced by cold, heat, or salt shock were enriched in three developmental synexpression groups. In addition, certain proteins belonging to the heat and salt shock stimulons were coregulated during development. Thus, stress regulatory systems controlling these stimulons were implicated as integral parts of the developmental program. This correlation suggested that thermal shock and salt shock stress response regulatory systems either allow the cell to adapt to stresses associated with development or directly control the developmental program.

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“…In liquid, as in solid medium, the growth of Streptomyces was reported to be multiphasic (15,52), starting with a phase of active growth that ends with a transitory growth arrest called either the "transition phase" (15) or the "diauxic lag" (32,50,51). Then growth resumes for a few hours before entry of the bacteria into stationary phase.…”

Section: Discussionmentioning

confidence: 99%

Phosphoinositides Are Involved in Control of the Glucose-Dependent Growth Resumption That Follows the Transition Phase inStreptomyces lividans

Chouayekh

Nothaft

Delaunay³

et al. 2007

J Bacteriol

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The interruption of the sblA gene of Streptomyces lividans was previously shown to lead to relief of glucose repression of the normally strongly glucose-repressed ␣-amylase gene. In addition to this relief, an early entry into stationary phase was observed when cells were grown in a minimal medium containing glucose as the main carbon source. In this study, we established that this mutant does not resume growth after the transition phase when cultured in the complex glucose-rich liquid medium R2YE and sporulates much earlier than the wild-type strain when plated on solid R2YE. These phenotypic differences, which were abolished when glucose was omitted from the R2YE medium, correlated with a reduced glucose uptake ability of the sblA mutant strain. sblA was shown to encode a bifunctional enzyme possessing phospholipase C-like and phosphoinositide phosphatase activities. The cleavage of phosphoinositides by SblA seems necessary to trigger the glucose-dependent renewed growth that follows the transition phase. The transient expression of sblA that takes place just before the transition phase is consistent with a regulatory role for this gene during the late stages of growth. The tight temporal control of sblA expression was shown to depend on two operator sites. One, located just upstream of the ؊35 promoter region, likely constitutes a repressor binding site. The other, located 170 bp downstream of the GTG sblA translational start codon, may be involved in the regulation of the degradation of the sblA transcript. This study suggests that phosphoinositides constitute important regulatory molecules in Streptomyces, as they do in eukaryotes.

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Identification of procaryotic developmental stages by statistical analyses of two‐dimensional gel patterns

Cited by 51 publications

References 16 publications

Morphological changes and proteome response of Corynebacterium glutamicum to a partial depletion of FtsI

Morphological changes and proteome response of Corynebacterium glutamicum to a partial depletion of FtsI

Developmental Control of Stress Stimulons in Streptomyces coelicolor Revealed by Statistical Analyses of Global Gene Expression Patterns

Phosphoinositides Are Involved in Control of the Glucose-Dependent Growth Resumption That Follows the Transition Phase inStreptomyces lividans

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