Space- and time-resolved protein dynamics in single bacterial cells observed on a chip

Greif, Dominik; Pobigaylo, Nataliya; Frage, Benjamin; Becker, Anke; Regtmeier, Jan; Anselmetti, Dario

doi:10.1016/j.jbiotec.2010.06.003

Cited by 12 publications

(8 citation statements)

References 48 publications

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“…Compared to the control, induced overexpression of ecpR1 reduced p gcrA - 122+3 - egfp and pP gcrA - 122+3 - egfp mediated fluorescence to 34% and 42%, respectively ( Fig 4A ). Furthermore, activity of a chromosomally integrated gcrA 3’- egfp translational fusion [ 46 ] was reduced to 75% in response to ecpR1 overexpression, validating the two-plasmid assay and confirming that posttranscriptional repression of EcpR1 results in reduction of GcrA protein level.…”

Section: Resultsmentioning

confidence: 63%

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A Stress-Induced Small RNA Modulates Alpha-Rhizobial Cell Cycle Progression

et al. 2015

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Mechanisms adjusting replication initiation and cell cycle progression in response to environmental conditions are crucial for microbial survival. Functional characterization of the trans-encoded small non-coding RNA (trans-sRNA) EcpR1 in the plant-symbiotic alpha-proteobacterium Sinorhizobium meliloti revealed a role of this class of riboregulators in modulation of cell cycle regulation. EcpR1 is broadly conserved in at least five families of the Rhizobiales and is predicted to form a stable structure with two defined stem-loop domains. In S. meliloti, this trans-sRNA is encoded downstream of the divK-pleD operon. ecpR1 belongs to the stringent response regulon, and its expression was induced by various stress factors and in stationary phase. Induced EcpR1 overproduction led to cell elongation and increased DNA content, while deletion of ecpR1 resulted in reduced competitiveness. Computationally predicted EcpR1 targets were enriched with cell cycle-related mRNAs. Post-transcriptional repression of the cell cycle key regulatory genes gcrA and dnaA mediated by mRNA base-pairing with the strongly conserved loop 1 of EcpR1 was experimentally confirmed by two-plasmid differential gene expression assays and compensatory changes in sRNA and mRNA. Evidence is presented for EcpR1 promoting RNase E-dependent degradation of the dnaA mRNA. We propose that EcpR1 contributes to modulation of cell cycle regulation under detrimental conditions.

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Section: Resultsmentioning

confidence: 63%

“…In the non-synchronized cultures, this might have been due to heterogeneous expression of such genes dependent on the cell cycle state as has been described for gcrA in S . meliloti and other cell cycle-dependent genes whose transcription varies during cell cycle progression [ 25 , 46 ].…”

Section: Discussionmentioning

confidence: 99%

A Stress-Induced Small RNA Modulates Alpha-Rhizobial Cell Cycle Progression

et al. 2015

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“…Although conspicuous organelles that distinguish between the two poles have not yet been identified in S. meliloti , time‐lapse microscopy allowed observation of PodJ1 targeting to the new cell pole. Furthermore, our results reinforce previous findings showing that each cell division in S. meliloti is asymmetric, producing two cells with distinct sizes and molecular compositions (Lam et al ., 2003; Hallez et al ., 2004; 2007; Greif et al ., 2010; Ratcliff and Denison, 2010; Brown et al ., 2012). Following cytokinesis, in the larger mother cell, PodJ1 localizes to the same side as it had previously appeared, the cell pole where new daughter cells bud off.…”

Section: Discussionmentioning

confidence: 99%

“…Many of the regulatory circuits that control organelle development and cell polarity in C. crescentus have been elucidated in detail and shown to involve the interplay of factors with specific subcellular locations (Laub et al ., 2007). Although S. meliloti and other alphaproteobacteria appear morphologically quite distinct, critical genetic circuits identified in C. crescentus are typically conserved across the phylogenetic group (Wright et al ., 1997; Barnett et al ., 2001; Brilli et al ., 2010; Schneiker‐Bekel et al ., 2011), and key proteins exhibit similar localization patterns (Lam et al ., 2003; Hallez et al ., 2007; Greif et al ., 2010).…”

Section: Introductionmentioning

confidence: 99%

The conserved polarity factor PodJ1 impacts multiple cell envelope‐associated functions in Sinorhizobium meliloti

Fields

Navarrete

Zare

et al. 2012

Molecular Microbiology

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Summary Although diminutive in size, bacteria possess highly diverse and spatially confined cellular structures. Two related alpha-proteobacteria, Sinorhizobium meliloti and Caulobacter crescentus, serve as models for investigating the genetic basis of morphologic variations. S. meliloti, a symbiont of leguminous plants, synthesizes multiple flagella and no prosthecae, whereas C. crescentus, a freshwater bacterium, has a single polar flagellum and stalk. The podJ gene, originally identified in C. crescentus for its role in polar organelle development, is split into two adjacent open reading frames, podJ1 and podJ2, in S. meliloti. Deletion of podJ1 interferes with flagellar motility, exopolysaccharide production, cell envelope integrity, cell division, and normal morphology, but not symbiosis. As in C. crescentus, the S. meliloti PodJ1 protein appears to act as a polarity beacon and localizes to the newer cell pole. Microarray analysis indicates that podJ1 affects the expression of at least 129 genes, the majority of which correspond to observed mutant phenotypes. Together, phenotypic characterization, microarray analysis, and suppressor identification suggest that PodJ1 controls a core set of conserved elements, including flagellar and pili genes, the signaling proteins PleC and DivK, and the transcriptional activator TacA, while alternate downstream targets have evolved to suit the distinct lifestyles of individual species.

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“…As in B. abortus, the PdhS-like histidine kinase CbrA of S. meliloti acts, together with DivJ, as a major kinase of DivK (Pini et al 2013) and localizes DivK∼P to the old pole (Greif et al 2010;Lam et al 2003;Sadowksi et al 2013). PodJ, which in C. crescentus positions PleC (Viollier et al 2002), localizes to the new pole of the predivisional cell.…”

Section: Ctra-dependent Cell Cycle Regulation In Polarly Growing Rhizmentioning

confidence: 99%

Generating asymmetry in a changing environment: cell cycle regulation in dimorphic alphaproteobacteria

Teeseling

Thanbichler

2020

Biological Chemistry

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While many bacteria divide by symmetric binary fission, some alphaproteobacteria have strikingly asymmetric cell cycles, producing offspring that differs significantly in their morphology and reproductive state. To establish this asymmetry, these species employ a complex cell cycle regulatory pathway based on two-component signaling cascades. At the center of this network is the essential DNA-binding response regulator CtrA, which acts as a transcription factor controlling numerous genes with cell cycle-relevant functions as well as a regulator of chromosome replication. The DNA-binding activity of CtrA is controlled at the level of both protein phosphorylation and stability, dependent on an intricate network of regulatory proteins, whose function is tightly coordinated in time and space. CtrA is differentially activated in the two (developing) offspring, thereby establishing distinct transcriptional programs that ultimately determine their distinct cell fates. Phase-separated polar microdomains of changing composition sequester proteins involved in the (in-)activation and degradation of CtrA specifically at each pole. In this review, we summarize the current knowledge of the CtrA pathway and discuss how it has evolved to regulate the cell cycle of morphologically distinct alphaproteobacteria.

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Space- and time-resolved protein dynamics in single bacterial cells observed on a chip

Cited by 12 publications

References 48 publications

A Stress-Induced Small RNA Modulates Alpha-Rhizobial Cell Cycle Progression

A Stress-Induced Small RNA Modulates Alpha-Rhizobial Cell Cycle Progression

The conserved polarity factor PodJ1 impacts multiple cell envelope‐associated functions in Sinorhizobium meliloti

Generating asymmetry in a changing environment: cell cycle regulation in dimorphic alphaproteobacteria

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