Directed polar secretion of protease from single cells of Vibrio cholerae via the type II secretion pathway

AbstractThis study reveals that it is possible to secrete truncated versions of outer membrane cytochromes into the culture supernatant and that these proteins can provide a basis for the export of heterologously produced proteins. Different soluble and truncated versions of the outer membrane cytochrome MtrF were analyzed for their suitability to be secreted. A protein version with a very short truncation of the N-terminus to remove the recognition sequence for the addition of a lipid anchor is secreted efficiently to the culture supernatant, and moreover this protein could be further truncated by a deletion of 160 amino acid and still is detectable in the supernatant. By coupling a cellulase to this soluble outer membrane cytochrome, the export efficiency was measured by means of relative cellulase activity. We conclude that outer membrane cytochromes of S. oneidensis can be applied as transporters for the export of target proteins into the medium using the type II secretion pathway.

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Expression of the endogenous type II secretion pathway in Escherichia coli leads to chitinase secretion

The EMBO Journal ◽

10.1093/emboj/19.24.6697 ◽

2000 ◽

Vol 19 (24) ◽

pp. 6697-6703 ◽

Cited By ~ 98

Author(s):

O. Francetic

Keyword(s):

Escherichia Coli ◽

Type Ii Secretion ◽

Type Ii ◽

Secretion Pathway

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Cyclic Di-GMP and VpsR Induce the Expression of Type II Secretion in Vibrio cholerae

Journal of Bacteriology ◽

10.1128/jb.00106-17 ◽

2017 ◽

Vol 199 (19) ◽

Cited By ~ 14

Author(s):

Rudolph E. Sloup ◽

Ashley E. Konal ◽

Geoffrey B. Severin ◽

Michelle L. Korir ◽

Mira M. Bagdasarian ◽

...

Keyword(s):

Vibrio Cholerae ◽

Protein Complexes ◽

Second Messenger ◽

Type Ii Secretion ◽

Type Ii ◽

Secretion Systems ◽

Content Type ◽

Severe Diarrhea ◽

Wide Range ◽

Genes Encoding

ABSTRACT Vibrio cholerae is a human pathogen that alternates between growth in environmental reservoirs and infection of human hosts, causing severe diarrhea. The second messenger cyclic di-GMP (c-di-GMP) mediates this transition by controlling a wide range of functions, such as biofilms, virulence, and motility. Here, we report that c-di-GMP induces expression of the extracellular protein secretion (eps) gene cluster, which encodes the type II secretion system (T2SS) in V. cholerae. Analysis of the eps genes confirmed the presence of two promoters located upstream of epsC, the first gene in the operon, one of which is induced by c-di-GMP. This induction is directly mediated by the c-di-GMP-binding transcriptional activator VpsR. Increased expression of the eps operon did not impact secretion of extracellular toxin or biofilm formation but did increase expression of the pseudopilin protein EpsG on the cell surface. IMPORTANCE Type II secretion systems (T2SSs) are the primary molecular machines by which Gram-negative bacteria secrete proteins and protein complexes that are folded and assembled in the periplasm. The substrates of T2SSs include extracellular factors, such as proteases and toxins. Here, we show that the widely conserved second messenger cyclic di-GMP (c-di-GMP) upregulates expression of the eps genes encoding the T2SS in the pathogen V. cholerae via the c-di-GMP-dependent transcription factor VpsR.

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Hexamers of the Type II Secretion ATPase GspE from Vibrio cholerae with Increased ATPase Activity

Structure ◽

10.1016/j.str.2013.06.027 ◽

2013 ◽

Vol 21 (9) ◽

pp. 1707-1717 ◽

Cited By ~ 43

Author(s):

Connie Lu ◽

Stewart Turley ◽

Samuel T. Marionni ◽

Young-Jun Park ◽

Kelly K. Lee ◽

...

Keyword(s):

Atpase Activity ◽

Vibrio Cholerae ◽

Type Ii Secretion ◽

Type Ii

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Docking and Assembly of the Type II Secretion Complex of Vibrio cholerae

Journal of Bacteriology ◽

10.1128/jb.01701-08 ◽

2009 ◽

Vol 191 (9) ◽

pp. 3149-3161 ◽

Cited By ~ 56

Author(s):

Suzanne R. Lybarger ◽

Tanya L. Johnson ◽

Miranda D. Gray ◽

Aleksandra E. Sikora ◽

Maria Sandkvist

Keyword(s):

Vibrio Cholerae ◽

Protein Interactions ◽

Fluorescent Protein ◽

Outer Membrane Proteins ◽

Subcellular Location ◽

Type Ii Secretion ◽

Type Ii ◽

Protein Fusion ◽

Green Fluorescent

ABSTRACT Secretion of cholera toxin and other virulence factors from Vibrio cholerae is mediated by the type II secretion (T2S) apparatus, a multiprotein complex composed of both inner and outer membrane proteins. To better understand the mechanism by which the T2S complex coordinates translocation of its substrates, we are examining the protein-protein interactions of its components, encoded by the extracellular protein secretion (eps) genes. In this study, we took a cell biological approach, observing the dynamics of fluorescently tagged EpsC and EpsM proteins in vivo. We report that the level and context of fluorescent protein fusion expression can have a bold effect on subcellular location and that chromosomal, intraoperon expression conditions are optimal for determining the intracellular locations of fusion proteins. Fluorescently tagged, chromosomally expressed EpsC and EpsM form discrete foci along the lengths of the cells, different from the polar localization for green fluorescent protein (GFP)-EpsM previously described, as the fusions are balanced with all their interacting partner proteins within the T2S complex. Additionally, we observed that fluorescent foci in both chromosomal GFP-EpsC- and GFP-EpsM-expressing strains disperse upon deletion of epsD, suggesting that EpsD is critical to the localization of EpsC and EpsM and perhaps their assembly into the T2S complex.

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