Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
AuthorsHamed, Mohamed Belal
van Mellaert, Lieve
Luzhetskyy, Andriy N
MetadataShow full item record
AbstractFluorescent proteins are a major cell biology tool to analyze protein sub-cellular topology. Here we have applied this technology to study protein secretion in the Gram-positive bacterium Streptomyces lividans TK24, a widely used host for heterologous protein secretion biotechnology. Green and monomeric red fluorescent proteins were fused behind Sec (SPSec) or Tat (SPTat) signal peptides to direct them through the respective export pathway. Significant secretion of fluorescent eGFP and mRFP was observed exclusively through the Tat and Sec pathways, respectively. Plasmid over-expression was compared to a chromosomally integrated spSec-mRFP gene to allow monitoring secretion under high and low level synthesis in various media. Fluorimetric detection of SPSec-mRFP recorded folded states, while immuno-staining detected even non-folded topological intermediates. Secretion of SPSec-mRFP is unexpectedly complex, is regulated independently of cell growth phase and is influenced by the growth regime. At low level synthesis, highly efficient secretion occurs until it is turned off and secretory preforms accumulate. At high level synthesis, the secretory pathway overflows and proteins are driven to folding and subsequent degradation. High-level synthesis of heterologous secretory proteins, whether secretion competent or not, has a drastic effect on the endogenous secretome, depending on their secretion efficiency. These findings lay the foundations of dissecting how protein targeting and secretion are regulated by the interplay between the metabolome, secretion factors and stress responses in the S. lividans model.
AffiliationHIPS, Helmholtz-Institut für Pharmazeutische Forschung Saarland, Universitätscampus E8.1 66123 Saarbrücken, Germany.
The following license files are associated with this item:
- Creative Commons
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-ShareAlike 4.0 International
- Modelling the metabolism of protein secretion through the Tat route in Streptomyces lividans.
- Authors: Valverde JR, Gullón S, Mellado RP
- Issue date: 2018 Jun 14
- Multi-Omics and Targeted Approaches to Determine the Role of Cellular Proteases in <i>Streptomyces</i> Protein Secretion.
- Authors: Busche T, Tsolis KC, Koepff J, Rebets Y, Rückert C, Hamed MB, Bleidt A, Wiechert W, Lopatniuk M, Yousra A, Anné J, Karamanou S, Oldiges M, Kalinowski J, Luzhetskyy A, Economou A
- Issue date: 2018
- Overproduction of a Model Sec- and Tat-Dependent Secretory Protein Elicits Different Cellular Responses in Streptomyces lividans.
- Authors: Gullón S, Marín S, Mellado RP
- Issue date: 2015
- Increase in xylanase production by Streptomyces lividans through simultaneous use of the Sec- and Tat-dependent protein export systems.
- Authors: Gauthier C, Li H, Morosoli R
- Issue date: 2005 Jun
- Comparison of the Sec and Tat secretion pathways for heterologous protein production by Streptomyces lividans.
- Authors: Schaerlaekens K, Lammertyn E, Geukens N, De Keersmaeker S, Anné J, Van Mellaert L
- Issue date: 2004 Sep 9