scholarly journals Identification of an Operon, Pil-Chp, That Controls Twitching Motility and Virulence in Xylella fastidiosa

2011 ◽  
Vol 24 (10) ◽  
pp. 1198-1206 ◽  
Author(s):  
Luciana Cursino ◽  
Cheryl D. Galvani ◽  
Dusit Athinuwat ◽  
Paulo A. Zaini ◽  
Yaxin Li ◽  
...  
Keyword(s):  
Xylella Fastidiosa ◽  
Severe Disease ◽  
Type Iv Pili ◽  
Plant Diseases ◽  
Twitching Motility ◽  
In Planta ◽  
Disease Manifestation ◽  
Phytopathogenic Bacterium ◽  
Type Iv ◽  
Genes Encoding

Xylella fastidiosa is an important phytopathogenic bacterium that causes many serious plant diseases, including Pierce's disease of grapevines. Disease manifestation by X. fastidiosa is associated with the expression of several factors, including the type IV pili that are required for twitching motility. We provide evidence that an operon, named Pil-Chp, with genes homologous to those found in chemotaxis systems, regulates twitching motility. Transposon insertion into the pilL gene of the operon resulted in loss of twitching motility (pilL is homologous to cheA genes encoding kinases). The X. fastidiosa mutant maintained the type IV pili, indicating that the disrupted pilL or downstream operon genes are involved in pili function, and not biogenesis. The mutated X. fastidiosa produced less biofilm than wild-type cells, indicating that the operon contributes to biofilm formation. Finally, in planta the mutant produced delayed and less severe disease, indicating that the Pil-Chp operon contributes to the virulence of X. fastidiosa, presumably through its role in twitching motility.

scholarly journals Upstream Migration of Xylella fastidiosa via Pilus-Driven Twitching Motility

2005 ◽  
Vol 187 (16) ◽  
pp. 5560-5567 ◽  
Author(s):  
Yizhi Meng ◽  
Yaxin Li ◽  
Cheryl D. Galvani ◽  
Guixia Hao ◽  
James N. Turner ◽  
...  
Keyword(s):  
Vascular System ◽  
Xylella Fastidiosa ◽  
Xylem Sap ◽  
Type Iv Pili ◽  
Type I ◽  
Upstream Migration ◽  
Twitching Motility ◽  
In Planta ◽  
Type Iv ◽  
Xylem Sap Flow

ABSTRACT Xylella fastidiosa is a xylem-limited nonflagellated bacterium that causes economically important diseases of plants by developing biofilms that block xylem sap flow. How the bacterium is translocated downward in the host plant's vascular system against the direction of the transpiration stream has long been a puzzling phenomenon. Using microfabricated chambers designed to mimic some of the features of xylem vessels, we discovered that X. fastidiosa migrates via type IV-pilus-mediated twitching motility at speeds up to 5 μm min−1 against a rapidly flowing medium (20,000 μm min−1). Electron microscopy revealed that there are two length classes of pili, long type IV pili (1.0 to 5.8 μm) and short type I pili (0.4 to 1.0 μm). We further demonstrated that two knockout mutants (pilB and pilQ mutants) that are deficient in type IV pili do not twitch and are inhibited from colonizing upstream vascular regions in planta. In addition, mutants with insertions in pilB or pilQ (possessing type I pili only) express enhanced biofilm formation, whereas a mutant with an insertion in fimA (possessing only type IV pili) is biofilm deficient.

scholarly journals Ralstonia solanacearum Needs Flp Pili for Virulence on Potato

2012 ◽  
Vol 25 (4) ◽  
pp. 546-556 ◽  
Author(s):  
Charles K. Wairuri ◽  
Jacquie E. van der Waals ◽  
Antoinette van Schalkwyk ◽  
Jacques Theron
Keyword(s):  
Ralstonia Solanacearum ◽  
Type Iv Pili ◽  
Periodontal Pathogen ◽  
Wild Type ◽  
In Planta ◽  
Phytopathogenic Bacterium ◽  
Type Iv ◽  
Plant Pathogenesis

Type IV pili are virulence factors in various bacteria. Several subclasses of type IV pili have been described according to the characteristics of the structural prepilin subunit. Although type IVa pili have been implicated in the virulence of Ralstonia solanacearum, type IVb pili have not previously been described in this plant pathogen. Here, we report the characterization of two distinct tad loci in the R. solanacearum genome. The tad genes encode functions necessary for biogenesis of the Flp subfamily of type IVb pili initially described for the periodontal pathogen Aggregatibacter actinomycetemcomitans. To determine the role of the tad loci in R. solanacearum virulence, we mutated the tadA2 gene located in the megaplasmid that encodes a predicted NTPase previously reported to function as the energizer for Flp pilus biogenesis. Characterization of the tadA2 mutant revealed that it was not growth impaired in vitro or in planta, produced wild-type levels of exopolysaccharide galactosamine, and exhibited swimming and twitching motility comparable with the wild-type strain. However, the tadA2 mutant was impaired in its ability to cause wilting of potato plants. This is the first report where type IVb pili in a phytopathogenic bacterium contribute significantly to plant pathogenesis.

scholarly journals Type I and type IV pili of Xylella fastidiosa affect twitching motility, biofilm formation and cell–cell aggregation

Microbiology ◽  
2007 ◽  
Vol 153 (3) ◽  
pp. 719-726 ◽  
Author(s):  
Yaxin. Li ◽  
Guixia. Hao ◽  
Cheryl D. Galvani ◽  
Yizhi Meng ◽  
Leonardo De La. Fuente ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Xylella Fastidiosa ◽  
Cell Aggregation ◽  
Type Iv Pili ◽  
Type I ◽  
Twitching Motility ◽  
Type Iv ◽  
Cell Cell

scholarly journals Mutations in Type I and Type IV Pilus Biosynthetic Genes Affect Twitching Motility Rates in Xylella fastidiosa

2007 ◽  
Vol 189 (20) ◽  
pp. 7507-7510 ◽  
Author(s):  
Leonardo De La Fuente ◽  
Thomas J. Burr ◽  
Harvey C. Hoch
Keyword(s):  
Standard Error ◽  
Flow Direction ◽  
Xylella Fastidiosa ◽  
Type Iv Pili ◽  
Type I ◽  
Twitching Motility ◽  
Wild Type ◽  
Type Iv Pilus ◽  
Type Iv ◽  
Rate Of Movement

ABSTRACT Xylella fastidiosa possesses both type I and type IV pili at the same cell pole. By use of a microfluidic device, the speed of twitching movement by wild-type cells on a glass surface against the flow direction of media was measured as 0.86 (standard error [SE], 0.04) μm min−1. A type I pilus mutant (fimA) moved six times faster (4.85 [SE, 0.27] μm min−1) and a pilY1 mutant moved three times slower (0.28 [SE, 0.03] μm min−1) than wild-type cells. Type I pili slow the rate of movement, while the putative type IV pilus protein PilY1 is likely important for attachment to surfaces.

Role of σ54 in the regulation of genes involved in type I and type IV pili biogenesis in Xylella fastidiosa

2007 ◽  
Vol 189 (3) ◽  
pp. 249-261 ◽  
Author(s):  
José F. da Silva Neto ◽  
Tie Koide ◽  
Cecília M. Abe ◽  
Suely L. Gomes ◽  
Marilis V. Marques
Keyword(s):  
Xylella Fastidiosa ◽  
Type Iv Pili ◽  
Type I ◽  
Type Iv

scholarly journals Motility and adhesion through type IV pili in Gram-positive bacteria

2016 ◽  
Vol 44 (6) ◽  
pp. 1659-1666 ◽  
Author(s):  
Kurt H. Piepenbrink ◽  
Eric J. Sundberg
Keyword(s):  
Bacterial Species ◽  
Cellular Adhesion ◽  
Type Iv Pili ◽  
Twitching Motility ◽  
Gram Positive ◽  
Bacterial Surface ◽  
Gram Positive Bacteria ◽  
Type Iv ◽  
Current State ◽  
Highly Hydrophobic

Type IV pili are hair-like bacterial surface appendages that play a role in diverse processes such as cellular adhesion, colonization, twitching motility, biofilm formation, and horizontal gene transfer. These extracellular fibers are composed exclusively or primarily of many copies of one or more pilin proteins, tightly packed in a helix so that the highly hydrophobic amino-terminus of the pilin is buried in the pilus core. Type IV pili have been characterized extensively in Gram-negative bacteria, and recent advances in high-throughput genomic sequencing have revealed that they are also widespread in Gram-positive bacteria. Here, we review the current state of knowledge of type IV pilus systems in Gram-positive bacterial species and discuss them in the broader context of eubacterial type IV pili.

scholarly journals Characterization of type IV pili in the life cycle of the predator bacterium Bdellovibrio

Microbiology ◽  
2010 ◽  
Vol 156 (4) ◽  
pp. 1040-1051 ◽  
Author(s):  
Khaled K. Mahmoud ◽  
Susan F. Koval
Keyword(s):  
Life Cycle ◽  
Insoluble Fraction ◽  
Type Iv Pili ◽  
Host Cells ◽  
Prey Cell ◽  
Gram Negative ◽  
Type Iv ◽  
Genes Encoding ◽  
Attack Phase

Bdellovibrio and like organisms (BALOs) are obligate prokaryotic predators of other Gram-negative bacteria. Bdellovibrio bacteriovorus is the most studied organism among BALOs. It has a periplasmic life cycle with two major stages: a motile, non-replicative stage spent searching for prey (the attack phase) and a stage spent inside the periplasm of the Gram-negative prey cell (the growth phase) after forming an osmotically stable body termed the bdelloplast. Within Bdellovibrio, there are also strains exhibiting an epibiotic life cycle. The genome sequence of the type strain B. bacteriovorus HD100T revealed the presence of multiple dispersed pil genes encoding type IV pili. Type IV pili in other bacteria are involved in adherence to and invasion of host cells and therefore can be considered to play a role in invasion of prey cells by Bdellovibrio. In this study, genes involved in producing type IV pili were identified in the periplasmic strain B. bacteriovorus 109J and an epibiotic Bdellovibrio sp. strain JSS. The presence of fibres on attack-phase cells was confirmed by examining negative stains of cells fixed with 10 % buffered formalin. Fibres were at the non-flagellated pole on approximately 25 % of attack-phase cells. To confirm that these fibres were type IV pili, a truncated form of PilA lacking the first 35 amino acids was designed to facilitate purification of the protein. The truncated PilA fused to a His-tag was overexpressed in Escherichia coli BL21(DE3) plysS. The fusion protein, accumulated in the insoluble fraction, was purified under denaturing conditions and used to produce polyclonal antisera. Immunoelectron microscopy showed that polar fibres present on the cell surface of the predator were composed of PilA, the major subunit of type IV pili. Immunofluorescence microscopy showed the presence of pilin on attack-phase cells of B. bacteriovorus 109J during attachment to prey cells and just after penetration, inside the bdelloplast. Antibodies against PilA delayed and inhibited predation in co-cultures of Bdellovibrio. This study confirms that type IV pili play a role in invasion of prey cells by Bdellovibrio.

scholarly journals Mucin inhibitsPseudomonas aeruginosabiofilm formation by significantly enhancing twitching motility

2014 ◽  
Vol 60 (3) ◽  
pp. 155-166 ◽  
Author(s):  
Cecily L. Haley ◽  
Cassandra Kruczek ◽  
Uzma Qaisar ◽  
Jane A. Colmer-Hamood ◽  
Abdul N. Hamood
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Type Iv Pili ◽  
Dependent Manner ◽  
Twitching Motility ◽  
Strain Pao1 ◽  
Concentration Dependent Manner ◽  
Dependent Effect ◽  
Type Iv ◽  
Biofilm Development

In Pseudomonas aeruginosa, type IV pili (TFP)-dependent twitching motility is required for development of surface-attached biofilm (SABF), yet excessive twitching motility is detrimental once SABF is established. In this study, we show that mucin significantly enhanced twitching motility and decreased SABF formation in strain PAO1 and other P. aeruginosa strains in a concentration-dependent manner. Mucin also disrupted partially established SABF. Our analyses revealed that mucin increased the amount of surface pilin and enhanced transcription of the pilin structural gene pilA. Mucin failed to enhance twitching motility in P. aeruginosa mutants defective in genes within the pilin biogenesis operons pilGHI/pilJK-chpA-E. Furthermore, mucin did not enhance twitching motility nor reduce biofilm development by chelating iron. We also examined the role of the virulence factor regulator Vfr in the effect of mucin. In the presence or absence of mucin, PAOΔvfr produced a significantly reduced SABF. However, mucin partially complemented the twitching motility defect of PAOΔvfr. These results suggest that mucin interferes with SABF formation at specific concentrations by enhancing TFP synthesis and twitching motility, that this effect, which is iron-independent, requires functional Vfr, and only part of the Vfr-dependent effect of mucin on SABF development occurs through twitching motility.

scholarly journals Expression of Type IV Pili by Moraxella catarrhalis Is Essential for Natural Competence and Is Affected by Iron Limitation

2004 ◽  
Vol 72 (11) ◽  
pp. 6262-6270 ◽  
Author(s):  
Nicole R. Luke ◽  
Amy J. Howlett ◽  
Jianqiang Shao ◽  
Anthony A. Campagnari
Keyword(s):  
Moraxella Catarrhalis ◽  
Pathogenic Bacteria ◽  
Type Iv Pili ◽  
Protein Subunit ◽  
Electron Microscopic ◽  
Type Iv ◽  
Wide Range ◽  
Genes Encoding ◽  
Microscopic Studies

ABSTRACT Type IV pili, filamentous surface appendages primarily composed of a single protein subunit termed pilin, play a crucial role in the initiation of disease by a wide range of pathogenic bacteria. Although previous electron microscopic studies suggested that pili might be present on the surface of Moraxella catarrhalis isolates, detailed molecular and phenotypic analyses of these structures have not been reported to date. We identified and cloned the M. catarrhalis genes encoding PilA, the major pilin subunit, PilQ, the outer membrane secretin through which the pilus filament is extruded, and PilT, the NTPase that mediates pilin disassembly and retraction. To initiate investigation of the role of this surface organelle in pathogenesis, isogenic pilA, pilT, and pilQ mutants were constructed in M. catarrhalis strain 7169. Comparative analyses of the wild-type 7169 strain and three isogenic pil mutants demonstrated that M. catarrhalis expresses type IV pili that are essential for natural genetic transformation. Our studies suggest type IV pilus production by M. catarrhalis is constitutive and ubiquitous, although pilin expression was demonstrated to be iron responsive and Fur regulated. These data indicate that additional studies aimed at elucidating the prevalence and role of type IV pili in the pathogenesis and host response to M. catarrhalis infections are warranted.

scholarly journals Calcium-Enhanced Twitching Motility in Xylella fastidiosa Is Linked to a Single PilY1 Homolog

2014 ◽  
Vol 80 (23) ◽  
pp. 7176-7185 ◽  
Author(s):  
Luisa F. Cruz ◽  
Jennifer K. Parker ◽  
Paul A. Cobine ◽  
Leonardo De La Fuente
Keyword(s):  
Xylella Fastidiosa ◽  
Xylem Sap ◽  
Active Movement ◽  
Binding Motif ◽  
Twitching Motility ◽  
Structural Protein ◽  
Plant Host ◽  
Content Type ◽  
Type Iv ◽  
Functional Differences

ABSTRACTThe plant-pathogenic bacteriumXylella fastidiosais restricted to the xylem vessel environment, where mineral nutrients are transported through the plant host; therefore, changes in the concentrations of these elements likely impact the growth and virulence of this bacterium. Twitching motility, dependent on type IV pili (TFP), is required for movement against the transpiration stream that results in basipetal colonization. We previously demonstrated that calcium (Ca) increases the motility ofX. fastidiosa, although the mechanism was unknown. PilY1 is a TFP structural protein recently shown to bind Ca and to regulate twitching and adhesion in bacterial pathogens of humans. Sequence analysis identified threepilY1homologs inX. fastidiosa(PD0023, PD0502, and PD1611), one of which (PD1611) contains a Ca-binding motif. Separate deletions of PD0023 and PD1611 resulted in mutants that still showed twitching motility and were not impaired in attachment or biofilm formation. However, the response of increased twitching at higher Ca concentrations was lost in thepilY1-1611 mutant. Ca does not modulate the expression of any of theX. fastidiosaPilY1 homologs, although it increases the expression of the retraction ATPasepilTduring active movement. The evidence presented here suggests functional differences between the PilY1 homologs, which may provideX. fastidiosawith an adaptive advantage in environments with high Ca concentrations, such as xylem sap.

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