Overproduction of the secretin OutD suppresses the secretion defect of an Erwinia chrysanthemi outB mutant

Condemine, Guy; Shevchik, Vladimir E.

doi:10.1099/00221287-146-3-639

Cited by 26 publications

(29 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recently, it was shown that two other Gsp proteins (probably) interact with the GspD secretin, i.e., XpsN (GspN) of Xanthomonas campestris (40) and OutB (GspB) of E. chrysanthemi (9). Genes encoding GspN have been identified in the gsp gene clusters of most GSP-containing bacteria, except E. chrysanthemi, P. aeruginosa, and P. alcaligenes (25,40).…”

Section: Discussionmentioning

confidence: 99%

Exchange of Xcp (Gsp) Secretion Machineries between Pseudomonas aeruginosa and Pseudomonas alcaligenes : Species Specificity Unrelated to Substrate Recognition

et al. 2001

View full text Add to dashboard Cite

Pseudomonas aeruginosa and Pseudomonas alcaligenes are gram-negative bacteria that secrete proteins using the type II or general secretory pathway, which requires at least 12 xcp gene products (XcpA and XcpP to -Z). Despite strong conservation of this secretion pathway, gram-negative bacteria usually cannot secrete exoproteins from other species. Based on results obtained with Erwinia, it has been proposed that the XcpP and/or XcpQ homologs determine this secretion specificity (M. Linderberg, G. P. Salmond, and A. Collmer, Mol. Microbiol. 20: [175][176][177][178][179][180][181][182][183][184][185][186][187][188][189][190] 1996). In the present study, we report that XcpP and XcpQ of P. alcaligenes could not substitute for their respective P. aeruginosa counterparts. However, these complementation failures could not be correlated to species-specific recognition of exoproteins, since these bacteria could secrete exoproteins of each other. Moreover, when P. alcaligenes xcpP and xcpQ were expressed simultaneously in a P. aeruginosa xcpPQ deletion mutant, complementation was observed, albeit only on agar plates and not in liquid cultures. After growth in liquid culture the heat-stable P. alcaligenes XcpQ multimers were not detected, whereas monomers were clearly visible. Together, our results indicate that the assembly of a functional Xcp machinery requires species-specific interactions between XcpP and XcpQ and between XcpP or XcpQ and another, as yet uncharacterized component(s).Many extracellular proteins produced by gram-negative bacteria are secreted by the type II secretion pathway, also called the main terminal branch (MTB) of the general secretory pathway (GSP) (49). Pseudomonas aeruginosa secretes several enzymes and toxins via this pathway (20). These proteins are translocated in two steps across the bacterial cell envelope.

show abstract

Section: Discussionmentioning

confidence: 99%

Exchange of Xcp (Gsp) Secretion Machineries between Pseudomonas aeruginosa and Pseudomonas alcaligenes : Species Specificity Unrelated to Substrate Recognition

et al. 2001

View full text Add to dashboard Cite

show abstract

“…One of the difficulties in determining whether PulS-like pilotins exist in other T2S systems is that the C-terminal regions of the secretins, to which they probably bind, are among the least conserved regions of these proteins, suggesting that the pilotins may also be poorly conserved. Other proteins, notably T2SA and T2SB (Ast et al, 2002;Condemine & Shevchik, 2000), have been proposed to perform PulS-like functions. Interestingly, E. coli K-12 has genes for T2SA and T2SB proteins within its secreton gene cluster (Francetic et al, 2000) as well as an unlinked gene, yacC, with limited similarity to pulS.…”

Section: T2s Systemsmentioning

confidence: 99%

Type II protein secretion and its relationship to bacterial type IV pili and archaeal flagella

et al. 2003

View full text Add to dashboard Cite

Homologues of the protein constituents of the Klebsiella pneumoniae (Klebsiella oxytoca) type II secreton (T2S), the Pseudomonas aeruginosa type IV pilus/fimbrium biogenesis machinery (T4P) and the Methanococcus voltae flagellum biogenesis machinery (Fla) have been identified. Known constituents of these systems include (1) a major prepilin (preflagellin), (2) several minor prepilins (preflagellins), (3) a prepilin (preflagellin) peptidase/methylase, (4) an ATPase, (5) a multispanning transmembrane (TM) protein, (6) an outer-membrane secretin (lacking in Fla) and (7) several functionally uncharacterized envelope proteins. Sequence and phylogenetic analyses led to the conclusion that, although many of the protein constituents are probably homologous, extensive sequence divergence during evolution clouds this homology so that a common ancestry can be established for all three types of systems for only two constituents, the ATPase and the TM protein. Sequence divergence of the individual T2S constituents has occurred at characteristic rates, apparently without shuffling of constituents between systems. The same is probably also true for the T4P and Fla systems. The family of ATPases is much larger than the family of TM proteins, and many ATPase homologues function in capacities unrelated to those considered here. Many phylogenetic clusters of the ATPases probably exhibit uniform function. Some of these have a corresponding TM protein homologue although others probably function without one. It is further shown that proteins that compose the different phylogenetic clusters in both the ATPase and the TM protein families exhibit unique structural characteristics that are of probable functional significance. The TM proteins are shown to have arisen by at least two dissimilar intragenic duplication events, one in the bacterial kingdom and one in the archaeal kingdom. The archaeal TM proteins are twice as large as the bacterial TM proteins, suggesting an oligomeric structure for the latter. OverviewThree related types of prokaryotic envelope protein complexes include (putative) prepilin proteins with highly similar hydrophobic N-terminal segments of approximately 20 amino acyl residues. These putative prepilins can assemble into filamentous structures which compose parts of (1) the type II secretion system (T2S), (2) the type IV piliation/fimbriation system (T4P) (both of Gramnegative bacteria) and (3) the flagellar system (Fla) of archaea. T2S, also called the type II secreton or the main terminal branch (MTB; TC #3.A.15) of the general secretory pathway (TC #3.A.5;Cao & Saier, 2003), represents the major pathway for exoprotein transport from the periplasm across the outer membrane in a wide variety of Gramnegative bacteria (Pugsley, 1993a). The type II secreton is composed of a core of around 12 proteins, some of which are not present in all type II secretons and others of which appear to be dispensable for secreton function Pugsley, 1993a;Sandkvist, 2001). In this review, secreton components will be referred to accordi...

show abstract

“…One possible role of ExeB may involve a direct interaction with ExeD, as OutB and OutD can be cross-linked with formaldehyde in E. coli cells (9). To date, A. hydrophila cross-linking studies have not identified such a complex.…”

Section: Vol 187 2005mentioning

confidence: 99%

“…Although normally silent, the secreton genes of the E. coli gsp system, including gspAB, were shown to be required for secretion of chitinase (13). The importance of GspB proteins in other translocation systems, such as the pul and out systems of Klebsiella and Erwinia, has also been characterized (8,9).…”

mentioning

confidence: 99%

Purification and Characterization of the N-Terminal Domain of ExeA: a Novel ATPase Involved in the Type II Secretion Pathway of Aeromonas hydrophila

Schoenhofen

Strozen

et al. 2005

J Bacteriol

View full text Add to dashboard Cite

Aeromonas hydrophila secretes a number of degradative enzymes and toxins into the external milieu via the type II secretory pathway or secreton. ExeA is an essential component of this system and is necessary for the localization and/or multimerization of the secretin ExeD. ExeA contains two sequence motifs characteristic of the Walker superfamily of ATPases. Previous examination of substitution derivatives altered in these motifs suggested that ATP binding or hydrolysis is required for ExeAB complex formation and subsequent secretion function. To directly examine ExeA function, the N-terminal cytoplasmic domain of ExeA with the addition of a C-terminal hexahistidine tag (cytExeA) was overproduced in Escherichia coli and purified by metal chelate affinity and anion-exchange chromatographic techniques. Purified preparations of cytExeA exhibited ATPase activity in the presence of several divalent cations, Mg 2؉ being the preferred cation, with an optimum reaction temperature of ϳ37 to 42°C and an optimum pH of 7 to 8. cytExeA exhibited an apparent K m for Mg-ATP of 0.22 mM and a V max of 0.72 nmol min ؊1 mg ؊1 of protein. cytExeA displayed low specificity for nucleoside triphosphate substrates and was significantly inhibited by F-type ATPase inhibitors. Gel filtration analyses of cytExeA, ExeA, and ExeAB indicated that ExeA dimerizes and forms a very large complex with ExeB. These findings support a model whereby ExeAB utilizes energy derived from ATP hydrolysis to facilitate the correct localization and multimerization of the ExeD secretin.

show abstract

Overproduction of the secretin OutD suppresses the secretion defect of an Erwinia chrysanthemi outB mutant

Cited by 26 publications

References 44 publications

Exchange of Xcp (Gsp) Secretion Machineries between Pseudomonas aeruginosa and Pseudomonas alcaligenes : Species Specificity Unrelated to Substrate Recognition

Exchange of Xcp (Gsp) Secretion Machineries between Pseudomonas aeruginosa and Pseudomonas alcaligenes : Species Specificity Unrelated to Substrate Recognition

Type II protein secretion and its relationship to bacterial type IV pili and archaeal flagella

Purification and Characterization of the N-Terminal Domain of ExeA: a Novel ATPase Involved in the Type II Secretion Pathway of Aeromonas hydrophila

Contact Info

Product

Resources

About