Transcriptional Analysis of the
 groES
 -
 groEL1
 ,
 groEL2
 , and
 dnaK
 genes in
 Corynebacterium glutamicum
 : Characterization of Heat Shock-Induced Promoters

Barreiro, Carlos; González-Lavado, Eva; Pátek, Miroslav; Martı́n, Juan-Francisco

doi:10.1128/jb.186.14.4813-4817.2004

Cited by 50 publications

(41 citation statements)

References 26 publications

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“…To confirm that the selected heat conditions based on transcriptional studies result in significant changes in the heat shock-induced proteins, anti-DnaK and anti-GroEL antibodies were used to probe total C. glutamicum proteins. Results showed that heating at 40°C for 60 min is adequate for observing the complete heat shock response in the proteome and correlates well with the changes observed at the transcriptional level (3).…”

Section: Resultsmentioning

confidence: 59%

“…Our results showed that two proteins, DnaK and GrpE, which are translated from the same transcripts (dnaK-grpEdnaJ-hspR and dnaK-grpE [3]), occur in very different amounts in the proteome (Fig. 1), indicating the existence of posttranscriptional controls affecting the synthesis or degradation of GrpE compared to that of DnaK.…”

Section: Discussionmentioning

confidence: 91%

“…There are two groEL genes in the genome of C. glutamicum, namely groEL1, which forms part of a bicistronic groES-groEL1 operon, and a separate one, groEL2, expressed as a monocistronic transcript (3).…”

Section: R E T R a C T E Dmentioning

confidence: 99%

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Heat Shock Proteome Analysis of Wild-Type Corynebacterium glutamicum ATCC 13032 and a Spontaneous Mutant Lacking GroEL1, a Dispensable Chaperone

et al. 2005

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Proteome analysis of Corynebacterium glutamicum ATCC 13032 showed that levels of several proteins increased drastically in response to heat shock. These proteins were identified as DnaK, GroEL1, GroEL2, ClpB, GrpE, and PoxB, and their heat response was in agreement with previous transcriptomic results. A major heat-induced protein was absent in the proteome of strain 13032B of C. glutamicum, used for genome sequencing in Germany, compared with the wild-type ATCC 13032 strain. The missing protein was identified as GroEL1 by matrix-assisted laser desorption ionization-time of flight peptide mass fingerprinting, and the mutation was found to be due to an insertion sequence, IsCg1, that was integrated at position 327 downstream of the translation start codon of the groEL1 gene, resulting in a truncated transcript of this gene, as shown by Northern analysis. The GroEL1 chaperone is, therefore, dispensable in C. glutamicum. On the other hand, GroEL2 appears to be essential for growth. Based on these results, the role of the duplicate groEL1 and groEL2 genes is analyzed.Corynebacterium glutamicum (initially named Micrococcus glutamicus) was isolated from a soil sample of the Ueno Zoo in Tokyo as a high producer of glutamic acid (22). The type strain Kyowa Hakko Kogyo strain 534 was reclassified as C. glutamicum (37) and deposited as ATCC 13032.A large number of mutants derived from C. glutamicum ATCC 13032 have been obtained in several laboratories and used for the industrial production of L-glutamic acid, L-lysine, and L-threonine. Due to its industrial relevance, a large research effort for the last two decades has focused on the molecular genetics of this bacterium (4,7,11,23,25,27). The genome of the Kyowa type strain ATCC 13032 was sequenced in Japan (19). In parallel, the genome of a different ATCC 13032 derivative (hereafter named C. glutamicum 13032B) was mapped (4) and sequenced in Germany (20).Heat shock of C. glutamicum cultures increases the secretion of glutamic acid (9) and lysine (30), and it favors the entry of exogenous DNA during transformation by electroporation (38). As in other microorganisms, heat treatment triggers a heat shock response in C. glutamicum (3,29).There is considerable interest in the analysis of the heat shock-induced proteins in C. glutamicum and the heat shock effect on (i) wide-domain regulatory mechanisms and (ii) specific enzymatic steps involved in biosynthesis (24) and secretion (6) of amino acids. The availability of the genome sequence and the improvement of the two-dimensional (2D) protein electrophoresis systems (17, 18, 35) have allowed good progress in the resolution and identification of many proteins of the C. glutamicum proteome. During a collaboration study on the proteome response to heat shock, we observed that a major heat-induced protein was absent in the C. glutamicum 13032B strain available in Bielefeld, Germany, which was present in the ATCC 13032 strain in our laboratory in León, Spain. It was therefore interesting to perform a detailed analysis of the heat ...

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Section: Resultsmentioning

confidence: 59%

Section: Discussionmentioning

confidence: 91%

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Heat Shock Proteome Analysis of Wild-Type Corynebacterium glutamicum ATCC 13032 and a Spontaneous Mutant Lacking GroEL1, a Dispensable Chaperone

et al. 2005

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“…Chaperonin proteins are found in almost all organisms (if not all) (Braig, 1998) and are classified into six families: Hsp10, Hsp40, Hsp60, Hsp70, Hsp90, and Hsp100. However, in prokaryotes, the HSPs most characterized are hsp60 (groEL), hsp70 (dnaK) and hsp10 (groES) (Eom et al, 2005), and in the Corynebacterium genus two paralogue genes of hsp60 have been described (Barreiro et al, 2004).…”

Section: Heat Shock Proteins: Chaperonin Immunogenicity and Vaccine Dmentioning

confidence: 99%

A description of genes of Corynebacterium pseudotuberculosis useful in diagnostics and vaccine applications

D’Afonseca

Moraes²,

Dorella³

et al. 2008

Genet. Mol. Res.

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ABSTRACT. Corynebacterium pseudotuberculosis, a Gram-positive intracellular pathogen, is the etiological agent of caseous lymphadenitis or CLA. This bacterium infects goats and sheep and causes great economic losses worldwide annually, mainly for goat producers. Despite its importance, CLA is still poorly characterized. However, with advances in the genomic field, many C. pseudotuberculosis genes have already been characterized, mainly those related to virulence such as phospholipase D. Here, we examined the use of the several available genes of C. pseudotuberculosis and reviewed their applications in vaccine construction, more efficient diagnostics for CLA, and control of this disease, among other applications.

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“…These genes are groES, groEL, dnaJ, grpE, and clpB, which encode chaperones and hspR, which encodes a transcriptional regulator of the heat shock response system (3,12,67). Moreover, four genes that encode hypothetical proteins, as well as a putative starvation-induced, DNA-protecting protein, were found to be differentially expressed (Table 1, genes with other functions).…”

Section: Resultsmentioning

confidence: 99%

The Extracytoplasmic Function-Type Sigma Factor SigM of Corynebacterium glutamicum ATCC 13032 Is Involved in Transcription of Disulfide Stress-Related Genes

et al. 2007

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The gene for the extracytoplasmic function (ECF) sigma factor SigM was deleted from the chromosome of the gram-positive soil bacterium Corynebacterium glutamicum to elucidate the role of the SigM protein in the regulation of gene expression. Comparative DNA microarray hybridizations of the C. glutamicum wild type and sigM-deficient mutant C. glutamicum DN1 revealed 23 genes with enhanced expression in the sigM-proficient strain, encoding functions in the assembly of iron-sulfur clusters (suf operon), thioredoxin reductase (trxB), thioredoxins (trxC, trxB1), chaperones (groES, groEL, clpB), and proteins involved in the heat shock response (hspR, dnaJ, grpE). Deletion of the sigM gene rendered the C. glutamicum cells more sensitive to heat, cold, and the presence of the thiol oxidant diamide. Transcription of the sigM gene increased under different stress conditions, including heat shock, cold shock, and disulfide stress caused by diamide treatment, suggesting a regulatory role for SigM under thiol-oxidative stress conditions. Stress-responsive promoters were determined upstream of the suf operon and of the trxB, trxC, and trxB1 genes. The deduced SigM consensus promoter is characterized by the ؊35 hexamer gGGAAT and the ؊10 hexamer YGTTGR. Transcription of the sigM gene is apparently controlled by the ECF sigma factor SigH, since a sigH mutant was unable to enhance the expression of sigM and the SigM regulon under thioloxidative stress conditions. A typical SigH-responsive promoter was mapped upstream of the sigM gene. The ECF sigma factor SigM is apparently part of a regulatory cascade, and its transcription is controlled by SigH under conditions of thiol-oxidative stress.Corynebacterium glutamicum is a gram-positive, nonsporulating soil bacterium that belongs to the order Actinomycetales, which also includes the genera Mycobacterium and Streptomyces. C. glutamicum is an industrially relevant producer of several L-amino acids (21, 40). Because of its biotechnological importance, the genetic and biochemical background of amino acid biosynthesis has been studied extensively. In recent years, these studies have been broadened by the completion of the genome sequence of the wild-type strain C. glutamicum ATCC 13032 (26, 30). The genome sequence allows the identification of genes by similarity to already known genes in other organisms. However, knowledge about gene expression and its regulation can only be derived from complementary studies. One attempt is based on the detection and classification of transcriptional regulators by bioinformatic methods (6), and the other is based on generating specific regulatory mutants and analyzing changes in gene expression by DNA microarray hybridization and real-time reverse transcription (RT)-PCR assays (25,54,36,7). It is also essential to store this information on transcriptional regulators and their regulatory interactions in the form of specialized databases such as CoryneRegNet, a data warehouse for corynebacterial transcription factors and gene regulatory networks (4). Mo...

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Transcriptional Analysis of the groES - groEL1 , groEL2 , and dnaK genes in Corynebacterium glutamicum : Characterization of Heat Shock-Induced Promoters

Cited by 50 publications

References 26 publications

Heat Shock Proteome Analysis of Wild-Type Corynebacterium glutamicum ATCC 13032 and a Spontaneous Mutant Lacking GroEL1, a Dispensable Chaperone

Heat Shock Proteome Analysis of Wild-Type Corynebacterium glutamicum ATCC 13032 and a Spontaneous Mutant Lacking GroEL1, a Dispensable Chaperone

A description of genes of Corynebacterium pseudotuberculosis useful in diagnostics and vaccine applications

The Extracytoplasmic Function-Type Sigma Factor SigM of Corynebacterium glutamicum ATCC 13032 Is Involved in Transcription of Disulfide Stress-Related Genes

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