Tetrameric c-di-GMP Mediates Effective Transcription Factor Dimerization to Control Streptomyces Development

Tschowri, Natalia; Schumacher, Maria A.; Schlimpert, Susan; Chinnam, Naga Babu; Findlay, Kim; Brennan, Richard G.; Buttner, Mark J.

doi:10.1016/j.cell.2014.07.022

Cited by 222 publications

(322 citation statements)

References 59 publications

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“…Binding of tetrameric CDG to BldD brings together the DNA binding domains of the BldD dimer, thus enabling the protein to bind to its target sites (224). An example of metabolic control is presented by the pleiotropic nutrient sensory regulator DasR, which is essential for development and pleiotropically represses antibiotic production (see below).…”

Section: The Streptomyces Life Cyclementioning

confidence: 99%

“…Recently, it was elegantly shown by the group of Mark Buttner that the activity of BldD, which represses many developmental genes during vegetative growth, is controlled posttranslationally by the signaling molecule cyclic-di-GMP (CDG) (224). Binding of tetrameric CDG to BldD brings together the DNA binding domains of the BldD dimer, thus enabling the protein to bind to its target sites (224).…”

Section: The Streptomyces Life Cyclementioning

confidence: 99%

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Taxonomy, Physiology, and Natural Products of Actinobacteria

Barka

Vatsa

Sanchez

et al. 2016

Microbiol Mol Biol Rev

1,524

1,048

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SUMMARYActinobacteriaare Gram-positive bacteria with high G+C DNA content that constitute one of the largest bacterial phyla, and they are ubiquitously distributed in both aquatic and terrestrial ecosystems. ManyActinobacteriahave a mycelial lifestyle and undergo complex morphological differentiation. They also have an extensive secondary metabolism and produce about two-thirds of all naturally derived antibiotics in current clinical use, as well as many anticancer, anthelmintic, and antifungal compounds. Consequently, these bacteria are of major importance for biotechnology, medicine, and agriculture.Actinobacteriaplay diverse roles in their associations with various higher organisms, since their members have adopted different lifestyles, and the phylum includes pathogens (notably, species ofCorynebacterium,Mycobacterium,Nocardia,Propionibacterium, andTropheryma), soil inhabitants (e.g.,MicromonosporaandStreptomycesspecies), plant commensals (e.g.,Frankiaspp.), and gastrointestinal commensals (Bifidobacteriumspp.).Actinobacteriaalso play an important role as symbionts and as pathogens in plant-associated microbial communities. This review presents an update on the biology of this important bacterial phylum.

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Section: The Streptomyces Life Cyclementioning

confidence: 99%

Section: The Streptomyces Life Cyclementioning

confidence: 99%

Taxonomy, Physiology, and Natural Products of Actinobacteria

Barka

Vatsa

Sanchez

et al. 2016

Microbiol Mol Biol Rev

1,524

1,048

View full text Add to dashboard Cite

show abstract

“…Although a diverse set of DNA-binding proteins have been experimentally identified as c-di-GMP-binding proteins, MrkH is thus far the only DNA-binding protein predicted to harbor a PilZ domain (18,(27)(28)(29)(30)(31). Data indicate that c-di-GMP binding to MrkH stimulates its ability to bind a consensus DNA sequence called the MrkH box, TATCAA, located upstream of the −35 site in the mrkABCDF promoter (6,8).…”

mentioning

confidence: 99%

“…In addition to biofilm formation, processes regulated by c-di-GMP include motility, virulence, and the cell cycle (15)(16)(17). Moreover, recent work has revealed that c-di-GMP serves as the switch that controls Streptomyces development and secondary metabolite production (18). Our understanding of the proteins and networks that drive c-di-GMP-dependent processes are, however, still in its infancy, largely because of the difficulty in identifying c-di-GMP-binding motifs within effector proteins.…”

mentioning

confidence: 99%

Structures of the activator of K. pneumonia biofilm formation, MrkH, indicates PilZ domains involved in c-di-GMP and DNA binding

Schumacher

Zhang

2016

Proc. Natl. Acad. Sci. U.S.A.

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The pathogenesis of Klebsiella pneumonia is linked to the bacteria’s ability to form biofilms. Mannose-resistant Klebsiella-like (Mrk) hemagglutinins are critical for K. pneumonia biofilm development, and the expression of the genes encoding these proteins is activated by a 3′,5′-cyclic diguanylic acid (c-di-GMP)–regulated transcription factor, MrkH. To gain insight into MrkH function, we performed structural and biochemical analyses. Data revealed MrkH to be a monomer with a two-domain architecture consisting of a PilZ C-domain connected to an N domain that unexpectedly also harbors a PilZ-like fold. Comparison of apo- and c-di-GMP–bound MrkH structures reveals a large 138° interdomain rotation that is induced by binding an intercalated c-di-GMP dimer. c-di-GMP interacts with PilZ C-domain motifs 1 and 2 (RxxxR and D/NxSxxG) and a newly described c-di-GMP–binding motif in the MrkH N domain. Strikingly, these c-di-GMP–binding motifs also stabilize an open state conformation in apo MrkH via contacts from the PilZ motif 1 to residues in the C-domain motif 2 and the c-di-GMP–binding N-domain motif. Use of the same regions in apo structure stabilization and c-di-GMP interaction allows distinction between the states. Indeed, domain reorientation by c-di-GMP complexation with MrkH, which leads to a highly compacted structure, suggests a mechanism by which the protein is activated to bind DNA. To our knowledge, MrkH represents the first instance of specific DNA binding mediated by PilZ domains. The MrkH structures also pave the way for the rational design of inhibitors that target K. pneumonia biofilm formation.

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“…By growing Streptomyces in a microfluidic device connected to a fluorescence widefield microscope (Figure 2), we are now able to monitor germination, vegetative growth and sporulation septation over a time period of up to 30 hr. This is greatly facilitated by the use of the new model organism Streptomyces venezuelae because it sporulates to near completion in submerged culture and thereby overcomes the limitation of the classical model species S. coelicolor, which sporulates only on solid media [16][17][18][19][20] . To help visualize vegetative growth and sporulation, we co-express fluorescently tagged versions of the cell polarity marker DivIVA and the key cell division protein FtsZ.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence Time-lapse Imaging of the Complete <em>S. venezuelae</em> Life Cycle Using a Microfluidic Device

Schlimpert

Flärdh

Buttner

2016

JoVE

Self Cite

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Live-cell imaging of biological processes at the single cell level has been instrumental to our current understanding of the subcellular organization of bacterial cells. However, the application of time-lapse microscopy to study the cell biological processes underpinning development in the sporulating filamentous bacteria Streptomyces has been hampered by technical difficulties.Here we present a protocol to overcome these limitations by growing the new model species, Streptomyces venezuelae, in a commercially available microfluidic device which is connected to an inverted fluorescence widefield microscope. Unlike the classical model species, Streptomyces coelicolor, S. venezuelae sporulates in liquid, allowing the application of microfluidic growth chambers to cultivate and microscopically monitor the cellular development and differentiation of S. venezuelae over long time periods. In addition to monitoring morphological changes, the spatio-temporal distribution of fluorescently labeled target proteins can also be visualized by time-lapse microscopy. Moreover, the microfluidic platform offers the experimental flexibility to exchange the culture medium, which is used in the detailed protocol to stimulate sporulation of S. venezuelae in the microfluidic chamber. Images of the entire S. venezuelae life cycle are acquired at specific intervals and processed in the open-source software Fiji to produce movies of the recorded time-series.

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Tetrameric c-di-GMP Mediates Effective Transcription Factor Dimerization to Control Streptomyces Development

Cited by 222 publications

References 59 publications

Taxonomy, Physiology, and Natural Products of Actinobacteria

Taxonomy, Physiology, and Natural Products of Actinobacteria

Structures of the activator of K. pneumonia biofilm formation, MrkH, indicates PilZ domains involved in c-di-GMP and DNA binding

Fluorescence Time-lapse Imaging of the Complete <em>S. venezuelae</em> Life Cycle Using a Microfluidic Device

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