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Comparative proteomic analysis of Lactobacillus plantarum WCFS1 and ΔctsR mutant strains under physiological and heat stress conditions.

Russo P, de la Luz Mohedano M, Capozzi V, de Palencia PF, López P, Spano G, Fiocco D - Int J Mol Sci (2012)

Bottom Line: The proteomic analysis of L. plantarum WCFS1 and ctsR mutant strains confirms at the translational level the CtsR-mediated regulation of some members of the Clp family, as well as the heat induction of typical stress response genes.Heat activation of the putative CtsR regulon genes at transcriptional and translational levels, in the ΔctsR mutant, suggests additional regulative mechanisms, as is the case of hsp1.Furthermore, isoforms of ClpE with different molecular mass were found, which might contribute to CtsR quality control.

View Article: PubMed Central - PubMed

Affiliation: Department of Agriculture, Food and Environment Sciences, Via Napoli 25, Foggia 71122, Italy; E-Mails: p.russo@unifg.it (P.R.); vittorio.capozzi@gmail.com (V.C.).

ABSTRACT
Among Gram-positive bacteria, CtsR (Class Three Stress gene Repressor) mainly regulates the expression of genes encoding the Clp ATPases and the ClpP protease. To gain a better understanding of the biological significance of the CtsR regulon in response to heat-shock conditions, we performed a global proteomic analysis of Lactobacillus plantarum WCFS1 and ΔctsR mutant strains under optimal or heat stress temperatures. Total protein extracts from bacterial cells were analyzed by two-dimensional gel fractionation. By comparing maps from different culture conditions and different L. plantarum strains, image analysis revealed 23 spots with altered levels of expression. The proteomic analysis of L. plantarum WCFS1 and ctsR mutant strains confirms at the translational level the CtsR-mediated regulation of some members of the Clp family, as well as the heat induction of typical stress response genes. Heat activation of the putative CtsR regulon genes at transcriptional and translational levels, in the ΔctsR mutant, suggests additional regulative mechanisms, as is the case of hsp1. Furthermore, isoforms of ClpE with different molecular mass were found, which might contribute to CtsR quality control. Our results could add new outlooks in order to determine the complex biological role of CtsR-mediated stress response in lactic acid bacteria.

No MeSH data available.


Related in: MedlinePlus

2-DE analysis of L. plantarum protein extracts. 2D-gel analysis of total protein extracts of L. plantarum WCFS1 wild type and ΔctsR mutant strains, from either unstressed cultures (A and B, respectively) or after exposure to 42 °C for 30 min (C and D, respectively). Peptides were separated by IEF in the pI range of 4.0 to 7.0 in the first dimension and by 12% SDS-PAGE in the second dimension. Resolved peptides were visualized by SYPRO Ruby staining. Differentially expressed spots are indicated.
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f1-ijms-13-10680: 2-DE analysis of L. plantarum protein extracts. 2D-gel analysis of total protein extracts of L. plantarum WCFS1 wild type and ΔctsR mutant strains, from either unstressed cultures (A and B, respectively) or after exposure to 42 °C for 30 min (C and D, respectively). Peptides were separated by IEF in the pI range of 4.0 to 7.0 in the first dimension and by 12% SDS-PAGE in the second dimension. Resolved peptides were visualized by SYPRO Ruby staining. Differentially expressed spots are indicated.

Mentions: Total protein extracts from L. plantarum WCFS1 wild type and ΔctsR mutant strains from unstressed or heat-treated cultures were analyzed by 2D-gel fractionation, using a linear pH gradient (pH 4.0–7.0) during electrofocusing. The second dimension exhibited a separation range from 10 to 250 kDa. Representative gels of the four experimental conditions are depicted in Figure 1.


Comparative proteomic analysis of Lactobacillus plantarum WCFS1 and ΔctsR mutant strains under physiological and heat stress conditions.

Russo P, de la Luz Mohedano M, Capozzi V, de Palencia PF, López P, Spano G, Fiocco D - Int J Mol Sci (2012)

2-DE analysis of L. plantarum protein extracts. 2D-gel analysis of total protein extracts of L. plantarum WCFS1 wild type and ΔctsR mutant strains, from either unstressed cultures (A and B, respectively) or after exposure to 42 °C for 30 min (C and D, respectively). Peptides were separated by IEF in the pI range of 4.0 to 7.0 in the first dimension and by 12% SDS-PAGE in the second dimension. Resolved peptides were visualized by SYPRO Ruby staining. Differentially expressed spots are indicated.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3472708&req=5

f1-ijms-13-10680: 2-DE analysis of L. plantarum protein extracts. 2D-gel analysis of total protein extracts of L. plantarum WCFS1 wild type and ΔctsR mutant strains, from either unstressed cultures (A and B, respectively) or after exposure to 42 °C for 30 min (C and D, respectively). Peptides were separated by IEF in the pI range of 4.0 to 7.0 in the first dimension and by 12% SDS-PAGE in the second dimension. Resolved peptides were visualized by SYPRO Ruby staining. Differentially expressed spots are indicated.
Mentions: Total protein extracts from L. plantarum WCFS1 wild type and ΔctsR mutant strains from unstressed or heat-treated cultures were analyzed by 2D-gel fractionation, using a linear pH gradient (pH 4.0–7.0) during electrofocusing. The second dimension exhibited a separation range from 10 to 250 kDa. Representative gels of the four experimental conditions are depicted in Figure 1.

Bottom Line: The proteomic analysis of L. plantarum WCFS1 and ctsR mutant strains confirms at the translational level the CtsR-mediated regulation of some members of the Clp family, as well as the heat induction of typical stress response genes.Heat activation of the putative CtsR regulon genes at transcriptional and translational levels, in the ΔctsR mutant, suggests additional regulative mechanisms, as is the case of hsp1.Furthermore, isoforms of ClpE with different molecular mass were found, which might contribute to CtsR quality control.

View Article: PubMed Central - PubMed

Affiliation: Department of Agriculture, Food and Environment Sciences, Via Napoli 25, Foggia 71122, Italy; E-Mails: p.russo@unifg.it (P.R.); vittorio.capozzi@gmail.com (V.C.).

ABSTRACT
Among Gram-positive bacteria, CtsR (Class Three Stress gene Repressor) mainly regulates the expression of genes encoding the Clp ATPases and the ClpP protease. To gain a better understanding of the biological significance of the CtsR regulon in response to heat-shock conditions, we performed a global proteomic analysis of Lactobacillus plantarum WCFS1 and ΔctsR mutant strains under optimal or heat stress temperatures. Total protein extracts from bacterial cells were analyzed by two-dimensional gel fractionation. By comparing maps from different culture conditions and different L. plantarum strains, image analysis revealed 23 spots with altered levels of expression. The proteomic analysis of L. plantarum WCFS1 and ctsR mutant strains confirms at the translational level the CtsR-mediated regulation of some members of the Clp family, as well as the heat induction of typical stress response genes. Heat activation of the putative CtsR regulon genes at transcriptional and translational levels, in the ΔctsR mutant, suggests additional regulative mechanisms, as is the case of hsp1. Furthermore, isoforms of ClpE with different molecular mass were found, which might contribute to CtsR quality control. Our results could add new outlooks in order to determine the complex biological role of CtsR-mediated stress response in lactic acid bacteria.

No MeSH data available.


Related in: MedlinePlus