Bacillus protein secretion: an unfolding story

Harwood, Colin R.; Cranenburgh, Rocky M.

doi:10.1016/j.tim.2007.12.001

Cited by 167 publications

(128 citation statements)

References 55 publications

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“…Combined genetic, proteomic, and bioinformatic analyses have revealed that the vast majority of proteins secreted by B. subtilis leave the cytoplasm in an unfolded state via the general secretion (Sec) pathway (47). Upon translocation, these proteins fold into their active and protease-resistant conformation (19). A limited number of proteins are secreted via the so-called twin-arginine (RR) (Tat) pathway, which, in contrast to the Sec pathway, can facilitate the transport of fully folded proteins (16,35,37,38,42,45,53).…”

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confidence: 99%

High-Salinity Growth Conditions Promote Tat-Independent Secretion of Tat Substrates in Bacillus subtilis

Ploeg

Monteferrante

Piersma

et al. 2012

Appl Environ Microbiol

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bThe Gram-positive bacterium Bacillus subtilis contains two Tat translocases, which can facilitate transport of folded proteins across the plasma membrane. Previous research has shown that Tat-dependent protein secretion in B. subtilis is a highly selective process and that heterologous proteins, such as the green fluorescent protein (GFP), are poor Tat substrates in this organism. Nevertheless, when expressed in Escherichia coli, both B. subtilis Tat translocases facilitated exclusively Tat-dependent export of folded GFP when the twin-arginine (RR) signal peptides of the E. coli AmiA, DmsA, or MdoD proteins were attached. Therefore, the present studies were aimed at determining whether the same RR signal peptide-GFP precursors would also be exported Tat dependently in B. subtilis. In addition, we investigated the secretion of GFP fused to the full-length YwbN protein, a strict Tat substrate in B. subtilis. Several investigated GFP fusion proteins were indeed secreted in B. subtilis, but this secretion was shown to be completely Tat independent. At high-salinity growth conditions, the Tat-independent secretion of GFP as directed by the RR signal peptides from the E. coli AmiA, DmsA, or MdoD proteins was significantly enhanced, and this effect was strongest in strains lacking the TatAy-TatCy translocase. This implies that high environmental salinity has a negative influence on the avoidance of Tat-independent secretion of AmiA-GFP, DmsA-GFP, and MdoD-GFP. We conclude that as-yet-unidentified control mechanisms reject the investigated GFP fusion proteins for translocation by the B. subtilis Tat machinery and, at the same time, set limits to their Tat-independent secretion, presumably via the Sec pathway.

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confidence: 99%

High-Salinity Growth Conditions Promote Tat-Independent Secretion of Tat Substrates in Bacillus subtilis

Ploeg

Monteferrante

Piersma

et al. 2012

Appl Environ Microbiol

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“…B. subtilis is highly appreciated for biotechnological applications because it has a large capacity to secrete high-quality proteins into the culture medium and because it has the status of generally recognized as safe (18,38,50). Furthermore, B. subtilis is amenable to genetic engineering, and many expression systems are available (2,16,31,40,43,44).…”

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Stress-Responsive Systems Set Specific Limits to the Overproduction of Membrane Proteins in Bacillus subtilis

Zweers

Wiegert

Dijl

2009

Appl Environ Microbiol

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Essential membrane proteins are generally recognized as relevant potential drug targets due to their exposed localization in the cell envelope. Unfortunately, high-level production of membrane proteins for functional and structural analyses is often problematic. This is mainly due to their high overall hydrophobicity. To develop new concepts for membrane protein overproduction, we investigated whether the biogenesis of overproduced membrane proteins is affected by stress response-related proteolytic systems in the membrane. For this purpose, the well-established expression host Bacillus subtilis was used to overproduce eight essential membrane proteins from B. subtilis and Staphylococcus aureus. The results show that the W regulon (responding to cell envelope perturbations) and the CssRS two-component regulatory system (responding to unfolded exported proteins) set critical limits to membrane protein production in large quantities. The identified sigW or cssRS mutant B. subtilis strains with significantly improved capacity for membrane protein production are interesting candidate expression hosts for fundamental research and biotechnological applications. Importantly, our results pinpoint the interdependent expression and function of membrane-associated proteases as key parameters in bacterial membrane protein production.

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“…Nearly all of the components of the Sec system identified in E. coli have also been identified in B. subtilis and are biochemically well-characterized (van Wely et al, 2001;Harwood et al, 2008). Among these components, the peripheral membrane protein, SecA is considered to play a pivotal role in secretion.…”

Section: Deletion Of the C-terminus Of Secamentioning

confidence: 99%