scholarly journals Characterization of the Exopolysaccharide Biosynthesis Pathway in Myxococcus xanthus

2020 ◽  
Vol 202 (19) ◽  
Author(s):  
María Pérez-Burgos ◽  
Inmaculada García-Romero ◽  
Jana Jung ◽  
Eugenia Schander ◽  
Miguel A. Valvano ◽  
...  
Keyword(s):  
Repeat Unit ◽  
Myxococcus Xanthus ◽  
Type Iv Pilus ◽  
Fruiting Body Formation ◽  
Content Type ◽  
Body Formation ◽  
Polysaccharide Biosynthesis ◽  
Type Iv ◽  
Biosynthetic Machinery ◽  
Dependent Pathway

ABSTRACT Myxococcus xanthus arranges into two morphologically distinct biofilms depending on its nutritional status, i.e., coordinately spreading colonies in the presence of nutrients and spore-filled fruiting bodies in the absence of nutrients. A secreted polysaccharide, referred to as exopolysaccharide (EPS), is a structural component of both biofilms and is also important for type IV pilus-dependent motility and fruiting body formation. Here, we characterize the biosynthetic machinery responsible for EPS biosynthesis using bioinformatics, genetics, heterologous expression, and biochemical experiments. We show that this machinery constitutes a Wzx/Wzy-dependent pathway dedicated to EPS biosynthesis. Our data support that EpsZ (MXAN_7415) is the polyisoprenyl-phosphate hexose-1-phosphate transferase responsible for the initiation of the repeat unit synthesis. Heterologous expression experiments support that EpsZ has galactose-1-P transferase activity. Moreover, MXAN_7416, renamed WzxEPS, and MXAN_7442, renamed WzyEPS, are the Wzx flippase and Wzy polymerase responsible for translocation and polymerization of the EPS repeat unit, respectively. In this pathway, EpsV (MXAN_7421) also is the polysaccharide copolymerase and EpsY (MXAN_7417) the outer membrane polysaccharide export (OPX) protein. Mutants with single in-frame deletions in the five corresponding genes had defects in type IV pilus-dependent motility and a conditional defect in fruiting body formation. Furthermore, all five mutants were deficient in type IV pilus formation, and genetic analyses suggest that EPS and/or the EPS biosynthetic machinery stimulates type IV pilus extension. Additionally, we identify a polysaccharide biosynthesis gene cluster, which together with an orphan gene encoding an OPX protein make up a complete Wzx/Wzy-dependent pathway for synthesis of an unknown polysaccharide. IMPORTANCE The secreted polysaccharide referred to as exopolysaccharide (EPS) has important functions in the social life cycle of M. xanthus; however, little is known about how EPS is synthesized. Here, we characterized the EPS biosynthetic machinery and showed that it makes up a Wzx/Wzy-dependent pathway for polysaccharide biosynthesis. Mutants lacking a component of this pathway had reduced type IV pilus-dependent motility and a conditional defect in development. These analyses also suggest that EPS and/or the EPS biosynthetic machinery is important for type IV pilus formation.

scholarly journals Characterization of the exopolysaccharide biosynthesis pathway in Myxococcus xanthus

2020 ◽  
Author(s):  
María Pérez-Burgos ◽  
Inmaculada García-Romero ◽  
Jana Jung ◽  
Eugenia Schander ◽  
Miguel A. Valvano ◽  
...  
Keyword(s):  
Fruiting Body ◽  
Repeat Unit ◽  
Myxococcus Xanthus ◽  
Type Iv Pili ◽  
Fruiting Body Formation ◽  
Body Formation ◽  
Polysaccharide Biosynthesis ◽  
Type Iv ◽  
Biosynthetic Machinery ◽  
Dependent Pathway

AbstractMyxococcus xanthus arranges into two morphologically distinct biofilms depending on its nutritional status, i.e. coordinately spreading colonies in the presence of nutrients and spore-filled fruiting bodies in the absence of nutrients. A secreted polysaccharide referred to as exopolysaccharide (EPS) is a structural component of both biofilms and is also important for type IV pili-dependent motility and fruiting body formation. Here, we characterize the biosynthetic machinery responsible for EPS biosynthesis using bioinformatics, genetics, heterologous expression, and biochemical experiments. We show that this machinery constitutes a Wzx/Wzy-dependent pathway dedicated to EPS biosynthesis. Our data support that EpsZ (MXAN_7415) is the polyisoprenyl-phosphate hexose-1-phosphate transferase responsible for initiation of the repeat unit synthesis. Heterologous expression experiments support that EpsZ has galactose-1-P transferase activity. Moreover, MXAN_7416, renamed WzxEPS, and MXAN_7442, renamed WzyEPS, are the Wzx flippase and Wzy polymerase responsible for translocation and polymerization of the EPS repeat unit, respectively. Also, in this pathway, EpsV (MXAN_7421) is the polysaccharide co-polymerase and EpsY (MXAN_7417) the outer membrane polysaccharide export (OPX) protein. Mutants with single in-frame deletions in the five corresponding genes had defects in type IV pili-dependent motility and a conditional defect in fruiting body formation. Furthermore, all five mutants were deficient in type IV pili formation and genetic analyses suggest that EPS and/or the EPS biosynthetic machinery stimulates type IV pili extension. Additionally, we identify a polysaccharide biosynthesis gene cluster, which together with an orphan gene encoding an OPX protein make up a complete Wzx/Wzy-dependent pathway for synthesis of an unknown polysaccharide.ImportanceThe secreted polysaccharide referred to as exopolysaccharide (EPS) has important functions in the social life cycle of M. xanthus; however, little is known about how EPS is synthesized. Here, we characterized the EPS biosynthetic machinery and show that it makes up a Wzx/Wzy-dependent pathway for polysaccharide biosynthesis. Mutants lacking a component of this pathway had reduced type IV pili-dependent motility and a conditional defect in development. Also, these analysis suggest that EPS and/or the EPS biosynthetic machinery is important for type IV pili formation.

scholarly journals Cyclic Di-GMP Regulates Type IV Pilus-Dependent Motility in Myxococcus xanthus

2015 ◽  
Vol 198 (1) ◽  
pp. 77-90 ◽  
Author(s):  
Dorota Skotnicka ◽  
Tobias Petters ◽  
Jan Heering ◽  
Michael Hoppert ◽  
Volkhard Kaever ◽  
...  
Keyword(s):  
Myxococcus Xanthus ◽  
Lifestyle Changes ◽  
Gliding Motility ◽  
Cyclase Activity ◽  
Diguanylate Cyclase ◽  
Type Iv Pilus ◽  
Cell Agglutination ◽  
Content Type ◽  
Type Iv ◽  
Altered Cell

ABSTRACTThe nucleotide-based second messenger bis-(3′-5′)-cyclic dimeric GMP (c-di-GMP) is involved in regulating a plethora of processes in bacteria that are typically associated with lifestyle changes.Myxococcus xanthusundergoes major lifestyle changes in response to nutrient availability, with the formation of spreading colonies in the presence of nutrients and spore-filled fruiting bodies in the absence of nutrients. Here, we investigated the function of c-di-GMP inM. xanthusand show that this bacterium synthesizes c-di-GMP during growth. Manipulation of the c-di-GMP level by expression of either an active, heterologous diguanylate cyclase or an active, heterologous phosphodiesterase correlated with defects in type IV pilus (T4P)-dependent motility, whereas gliding motility was unaffected. An increased level of c-di-GMP correlated with reduced transcription of thepilAgene (which encodes the major pilin of T4P), reduced the assembly of T4P, and altered cell agglutination, whereas a decreased c-di-GMP level correlated with altered cell agglutination. The systematic inactivation of the 24 genes inM. xanthusencoding proteins containing GGDEF, EAL, or HD-GYP domains, which are associated with c-di-GMP synthesis, degradation, or binding, identified three genes encoding proteins important for T4P-dependent motility, whereas all mutants had normal gliding motility. Purified DmxA had diguanylate cyclase activity, whereas the hybrid histidine protein kinases TmoK and SgmT, each of which contains a GGDEF domain, did not have diguanylate cyclase activity. These results demonstrate that c-di-GMP is important for T4P-dependent motility inM. xanthus.IMPORTANCEWe provide the first direct evidence thatM. xanthussynthesizes c-di-GMP and demonstrate that c-di-GMP is important for T4P-dependent motility, whereas we did not obtain evidence that c-di-GMP regulates gliding motility. The data presented uncovered a novel mechanism for regulation of T4P-dependent motility, in which increased levels of c-di-GMP inhibit transcription of thepilAgene (which encodes the major pilin of T4P), ultimately resulting in the reduced assembly of T4P. Moreover, we identified an enzymatically active diguanylate cyclase that is important for T4P-dependent motility.

scholarly journals Conditional requirement of SglT for type IV pili function and S-motility in Myxococcus xanthus

Microbiology ◽  
2020 ◽  
Vol 166 (4) ◽  
pp. 349-358
Author(s):  
Vera Troselj ◽  
Darshankumar T. Pathak ◽  
Daniel Wall
Keyword(s):  
Myxococcus Xanthus ◽  
Critical Role ◽  
Point Mutations ◽  
Solid Surfaces ◽  
Temperature Sensitive ◽  
Fruiting Body Formation ◽  
Temperature Growth ◽  
Content Type ◽  
New Class ◽  
Type Iv

Myxobacteria exhibit complex social behaviors such as predation, outer membrane exchange and fruiting body formation. These behaviors depend on coordinated movements of cells on solid surfaces that involve social (S) motility. S-motility is powered by extension-retraction cycles of type 4 pili (Tfp) and exopolysaccharides (EPS) that provide a matrix for group cellular movement. Here, we characterized a new class of S-motility mutants in Myxococcus xanthus . These mutants have a distinctive phenotype: they lack S-motility even though they produce pili and EPS and the phenotype is temperature-sensitive. The point mutations were mapped to a single locus, MXAN_3284, named sglT. Similar to pilT mutants, sglT mutants are hyperpiliated and, strikingly, the temperature-sensitive phenotype is caused by null mutations. Our results indicate that SglT plays a critical role in Tfp function associated with pilus retraction and that the block in pili retraction is caused by a Tfp assembly defect in the absence of SglT at high-temperature growth.

scholarly journals FibA and PilA act cooperatively during fruiting body formation of Myxococcus xanthus

2006 ◽  
Vol 61 (5) ◽  
pp. 1283-1293 ◽  
Author(s):  
Pamela J. Bonner ◽  
Wesley P. Black ◽  
Zhaomin Yang ◽  
Lawrence J. Shimkets
Keyword(s):  
Fruiting Body ◽  
Myxococcus Xanthus ◽  
Fruiting Body Formation ◽  
Body Formation

Identification and initial characterization of a new pair of sibling sRNAs of Neisseria gonorrhoeae involved in type IV pilus biogenesis

Microbiology ◽  
2021 ◽  
Vol 167 (9) ◽  
Author(s):  
Marie Zachary ◽  
Susanne Bauer ◽  
Maximilian Klepsch ◽  
Katharina Wagler ◽  
Bruno Hüttel ◽  
...  
Keyword(s):  
Target Genes ◽  
Target Prediction ◽  
Type Iv Pilus ◽  
Content Type ◽  
Type Iv ◽  
Gene Expression Control ◽  
Initial Characterization ◽  
Pilus Biogenesis ◽  
Regulatory Rnas

Non-coding regulatory RNAs mediate post-transcriptional gene expression control by a variety of mechanisms relying mostly on base-pairing interactions with a target mRNA. Though a plethora of putative non-coding regulatory RNAs have been identified by global transcriptome analysis, knowledge about riboregulation in the pathogenic Neisseriae is still limited. Here we report the initial characterization of a pair of sRNAs of N. gonorrhoeae , TfpR1 and TfpR2, which exhibit a similar secondary structure and identical single-stranded seed regions, and therefore might be considered as sibling sRNAs. By combination of in silico target prediction and sRNA pulse expression followed by differential RNA sequencing we identified target genes of TfpR1 which are involved in type IV pilus biogenesis and DNA damage repair. We provide evidence that members of the TfpR1 regulon can also be targeted by the sibling TfpR2.

scholarly journals Multicellular Development in Myxococcus xanthus Is Stimulated by Predator-Prey Interactions

2007 ◽  
Vol 189 (15) ◽  
pp. 5675-5682 ◽  
Author(s):  
James E. Berleman ◽  
John R. Kirby
Keyword(s):  
Fruiting Body ◽  
Prey Availability ◽  
Process Analysis ◽  
Myxococcus Xanthus ◽  
Stringent Response ◽  
Fruiting Bodies ◽  
Fruiting Body Formation ◽  
Multicellular Development ◽  
Body Formation ◽  
Body Development

ABSTRACT Myxococcus xanthus is a predatory bacterium that exhibits complex social behavior. The most pronounced behavior is the aggregation of cells into raised fruiting body structures in which cells differentiate into stress-resistant spores. In the laboratory, monocultures of M. xanthus at a very high density will reproducibly induce hundreds of randomly localized fruiting bodies when exposed to low nutrient availability and a solid surface. In this report, we analyze how M. xanthus fruiting body development proceeds in a coculture with suitable prey. Our analysis indicates that when prey bacteria are provided as a nutrient source, fruiting body aggregation is more organized, such that fruiting bodies form specifically after a step-down or loss of prey availability, whereas a step-up in prey availability inhibits fruiting body formation. This localization of aggregates occurs independently of the basal nutrient levels tested, indicating that starvation is not required for this process. Analysis of early developmental signaling relA and asgD mutants indicates that they are capable of forming fruiting body aggregates in the presence of prey, demonstrating that the stringent response and A-signal production are surprisingly not required for the initiation of fruiting behavior. However, these strains are still defective in differentiating to spores. We conclude that fruiting body formation does not occur exclusively in response to starvation and propose an alternative model in which multicellular development is driven by the interactions between M. xanthus cells and their cognate prey.

scholarly journals Self-Driven Phase Transitions Drive Myxococcus xanthus Fruiting Body Formation

2019 ◽  
Vol 122 (24) ◽  
Author(s):  
Guannan Liu ◽  
Adam Patch ◽  
Fatmagül Bahar ◽  
David Yllanes ◽  
Roy D. Welch ◽  
...  
Keyword(s):  
Phase Transitions ◽  
Fruiting Body ◽  
Myxococcus Xanthus ◽  
Fruiting Body Formation ◽  
Body Formation

scholarly journals Gliding Motility Revisited: How Do the Myxobacteria Move without Flagella?

2010 ◽  
Vol 74 (2) ◽  
pp. 229-249 ◽  
Author(s):  
Emilia M. F. Mauriello ◽  
Tâm Mignot ◽  
Zhaomin Yang ◽  
David R. Zusman
Keyword(s):  
Biofilm Formation ◽  
Protein Localization ◽  
Myxococcus Xanthus ◽  
Gliding Motility ◽  
Solid Surfaces ◽  
Biological Functions ◽  
Fruiting Body Formation ◽  
Body Formation ◽  
Periplasmic Flagella ◽  
Motile Bacterium

SUMMARY In bacteria, motility is important for a wide variety of biological functions such as virulence, fruiting body formation, and biofilm formation. While most bacteria move by using specialized appendages, usually external or periplasmic flagella, some bacteria use other mechanisms for their movements that are less well characterized. These mechanisms do not always exhibit obvious motility structures. Myxococcus xanthus is a motile bacterium that does not produce flagella but glides slowly over solid surfaces. How M. xanthus moves has remained a puzzle that has challenged microbiologists for over 50 years. Fortunately, recent advances in the analysis of motility mutants, bioinformatics, and protein localization have revealed likely mechanisms for the two M. xanthus motility systems. These results are summarized in this review.

scholarly journals The Che4 pathway of Myxococcus xanthus regulates type IV pilus-mediated motility

2004 ◽  
Vol 52 (6) ◽  
pp. 1799-1811 ◽  
Author(s):  
Hera C. Vlamakis ◽  
John R. Kirby ◽  
David R. Zusman
Keyword(s):  
Myxococcus Xanthus ◽  
Type Iv Pilus ◽  
Type Iv
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