Clostridial DivIVA and MinD interact in the absence of MinJ

Valenčíková, Romana; Krascsenitsová, Eva; Labajová, Naďa; Makroczyová, Jana; Barák, Imrich

doi:10.1016/j.anaerobe.2018.01.013

Cited by 10 publications

(9 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…7), which is consistent with the B. subtilis model of cells committing to sporulation (33). The Clostridium divisome has not been well characterized due to the limitation of traditional FPs for viewing anaerobic cell division (40). It has been shown that the cell divisionassociated Min homolog in anaerobic Clostridioides difficile shows an oscillating pattern when tagged with yellow fluorescent protein (YFP) and expressed in B. subtilis (41).…”

Section: Discussionsupporting

confidence: 68%

“…It has been shown that the cell divisionassociated Min homolog in anaerobic Clostridioides difficile shows an oscillating pattern when tagged with yellow fluorescent protein (YFP) and expressed in B. subtilis (41). It was also shown that the DivIVA and MinD proteins show direct interaction in both C. difficile and Clostridium beijerinckii (40). The FAST-ZapA data presented here suggest the exciting potential of using FAST to tag other cell division proteins for live cell division imaging in C. acetobutylicum, other Clostridium spp., and, more broadly, other anaerobic prokaryotes.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

A Strongly Fluorescing Anaerobic Reporter and Protein-Tagging System for Clostridium Organisms Based on the Fluorescence-Activating and Absorption-Shifting Tag Protein (FAST)

Streett

Kalis

Papoutsakis

2019

Appl Environ Microbiol

View full text Add to dashboard Cite

Visualizing protein localization and characterizing gene expression activity in live Clostridium cells is limited for lack of a real-time, highly fluorescent, oxygen-independent reporter system. Enzymatic reporter systems have been used successfully for many years with Clostridium spp.; however, these assays do not allow for real-time analysis of gene expression activity with flow cytometry or for visualizing protein localization through fusion proteins. Commonly used fluorescent reporter proteins require oxygen for chromophore maturation and cannot be used for most strictly anaerobic Clostridium organisms. Here we show that the fluorescenceactivating and absorption-shifting tag protein (FAST), when associated with the fluorogenic ligand 4-hydroxy-3-methylbenzylidene-rhodanine (HMBR; now commercially available) and other commercially available ligands, is highly fluorescent in Clostridium acetobutylicum under anaerobic conditions. Using flow cytometry and a fluorescence microplate reader, we demonstrated FAST as a reporter system by employing the promoters of the C. acetobutylicum thiolase (thl), acetoacetate decarboxylase (adc), and phosphotransbutyrylase (ptb) metabolic genes, as well as a mutant P thl and modified ribosome binding site (RBS) versions of P adc and P ptb . Flow cytometrybased sorting was efficient and fast in sorting FAST-expressing cells, and positively and negatively sorted cells could be effectively recultured. FAST was also used to tag and examine protein localization of the predicted cell division FtsZ partner protein, ZapA, to visualize the divisome localization in live C. acetobutylicum cells. Our findings suggest that FAST can be used to further investigate Clostridium divisomes and more broadly the localization and expression levels of other proteins in Clostridium organisms, thus enabling cell biology studies with these organisms. IMPORTANCE FAST in association with the fluorogenic ligand HMBR is characterized as a successful, highly fluorescent reporter system in C. acetobutylicum. FAST can be used to distinguish between promoters in live cells using flow cytometry or a fluorescence microplate reader and can be used to tag and examine protein localization in live, anaerobically grown cells. Given that FAST is highly fluorescent under anaerobic conditions, it can be used in several applications of this and likely many Clostridium organisms and other strict anaerobes, including studies involving cell sorting, sporulation dynamics, and population characterization in pure as well as mixed cultures, such as those in various native or synthetic microbiomes and syntrophic cultures. Citation Streett HE, Kalis KM, Papoutsakis ET. 2019. A strongly fluorescing anaerobic reporter and protein-tagging system for Clostridium organisms based on the fluorescenceactivating and absorption-shifting tag protein (FAST). Appl Environ Microbiol 85:e00622-19. by guest http://aem.asm.org/ Downloaded from RESULTS Expression of FAST off the native and modified C. acetobutylicum thiolase promoter in E. coli and C...

show abstract

Section: Discussionsupporting

confidence: 68%

Section: Discussionmentioning

confidence: 99%

A Strongly Fluorescing Anaerobic Reporter and Protein-Tagging System for Clostridium Organisms Based on the Fluorescence-Activating and Absorption-Shifting Tag Protein (FAST)

Streett

Kalis

Papoutsakis

2019

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…subtilis (MinJ) and E . coli (MinE) proteins and that they evolved their own mechanism of Min and DivIVA interactions 79 . Whereas we have not found a minJ homologue in the C .…”

Section: Resultsmentioning

confidence: 99%

“…Liu et al . 79 described transcription of granulose-involved genes in C . acetobutylicum ATCC 824 but no similarity either in operon organization or in individual genes homologies was found with C .…”

Section: Resultsmentioning

confidence: 99%

Acidogenesis, solventogenesis, metabolic stress response and life cycle changes in Clostridium beijerinckii NRRL B-598 at the transcriptomic level

Patáková

Branská

Sedlář

et al. 2019

Sci Rep

View full text Add to dashboard Cite

Clostridium beijerinckii NRRL B-598 is a sporulating, butanol and hydrogen producing strain that utilizes carbohydrates by the acetone-butanol-ethanol (ABE) fermentative pathway. The pathway consists of two metabolic phases, acidogenesis and solventogenesis, from which the latter one can be coupled with sporulation. Thorough transcriptomic profiling during a complete life cycle and both metabolic phases completed with flow cytometry, microscopy and a metabolites analysis helped to find out key genes involved in particular cellular events. The description of genes/operons that are closely involved in metabolism or the cell cycle is a necessary condition for metabolic engineering of the strain and will be valuable for all C. beijerinckii strains and other Clostridial species. The study focused on glucose transport and catabolism, hydrogen formation, metabolic stress response, binary fission, motility/chemotaxis and sporulation, which resulted in the composition of the unique image reflecting clostridial population changes. Surprisingly, the main change in expression of individual genes was coupled with the sporulation start and not with the transition from acidogenic to solventogenic metabolism. As expected, solvents formation started at pH decrease and the accumulation of butyric and acetic acids in the cultivation medium.

show abstract

“…DivIVA also appears to play a role in cell division in L. monocytogenes by directly interacting with MinD, thereby circumventing the need for MinJ (93); the direct MinD-DivIVA interaction has also been documented in Clostridium spp. (94).…”

mentioning

confidence: 99%

¡vIVA la DivIVA!

2019

View full text Add to dashboard Cite

Reproduction in the bacterial kingdom predominantly occurs through binary fission—a process in which one parental cell is divided into two similarly sized daughter cells. How cell division, in conjunction with cell elongation and chromosome segregation, is orchestrated by a multitude of proteins has been an active area of research spanning the past few decades. Together, the monumental endeavors of multiple laboratories have identified several cell division and cell shape regulators as well as their underlying regulatory mechanisms in rod-shaped Escherichia coli and Bacillus subtilis, which serve as model organisms for Gram-negative and Gram-positive bacteria, respectively. Yet our understanding of bacterial cell division and morphology regulation is far from complete, especially in noncanonical and non-rod-shaped organisms. In this review, we focus on two proteins that are highly conserved in Gram-positive organisms, DivIVA and its homolog GpsB, and attempt to summarize the recent advances in this area of research and discuss their various roles in cell division, cell growth, and chromosome segregation in addition to their interactome and posttranslational regulation.

show abstract

Clostridial DivIVA and MinD interact in the absence of MinJ

Cited by 10 publications

References 41 publications

A Strongly Fluorescing Anaerobic Reporter and Protein-Tagging System for Clostridium Organisms Based on the Fluorescence-Activating and Absorption-Shifting Tag Protein (FAST)

A Strongly Fluorescing Anaerobic Reporter and Protein-Tagging System for Clostridium Organisms Based on the Fluorescence-Activating and Absorption-Shifting Tag Protein (FAST)

Acidogenesis, solventogenesis, metabolic stress response and life cycle changes in Clostridium beijerinckii NRRL B-598 at the transcriptomic level

¡vIVA la DivIVA!

Contact Info

Product

Resources

About