Nucleotide sequence of aBacillus subtilisgene homologous to thegrpEgene ofE. colilocated immediately upstream of thednaKgene

Wetzstein, Michael; Schumann, Wolfgang

doi:10.1093/nar/18.5.1289

Cited by 25 publications

(12 citation statements)

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“…The coding region would encode a protein whose amino acid sequence has identities and conservative substitutions with regard to bacterial homologs that fall within the range observed among the latter (6) and between them and a recently described homolog from the mitochondria of Saccharomyces cerevisiae (2,11,17) (data not shown). The overall organization of the grpE chromosomal region in M. mazei is similar to those of B. subtilis (39,40), Clostridium acetobutylicum (26), Borrelia burgdorferi (38), and Staphylococcus aureus (27). In these bacteria and in M. mazei, grpE is clustered with dnaK and dnaJ, located upstream of the former gene and preceded by another gene whose role is still incompletely understood.…”

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confidence: 92%

Archaeal grpE: transcription in two different morphologic stages of Methanosarcina mazei and comparison with dnaK and dnaJ

1995

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Transcription of the heat shock gene grpE was studied in two different morphologic stages of the archaeon Methanosarcina mazei S-6 that differ in resistance to physical and chemical traumas: single cells and packets. While single cells are directly exposed to environmental changes, such as temperature elevations, cells in packets are surrounded by intercellular and peripheral material that keeps them together in a globular structure which can reach several millimeters in diameter. grpE transcript levels determined by Northern (RNA) blotting peaked after a 15-min heat shock in single cells. In contrast, the highest transcript levels in packets were observed after the longest heat shock tested, 60 min. The same response profiles were demonstrated by primer extension experiments and S1 nuclease analysis. A comparison of the grpE response to heat shock with those of dnaK and dnaJ showed that the grpE transcript level was the most increased, closely followed by that of the dnaK transcript, with that of the dnaJ gene being the least augmented. Transcription of grpE started at the same site under normal and heat shock temperatures, and the transcript was consistently ϳ700 bases long. Codon usage patterns revealed that the three archaeal genes use most codons and have the same codon preference for 61% of the amino acids.A gene encoding a GrpE homolog has recently been cloned from the genome of the archaeon Methanosarcina mazei S-6 and sequenced (6). It is the first example of a grpE heat shock gene found within the domain Archaea. The Archaea are organisms phylogenetically distinct from Bacteria (eubacteria) and Eucarya (eucaryotes) (41). However, Archaea resemble eucaryotes in several molecular biological features more than they resemble bacteria and are closer to eucaryotes than bacteria (15,28,30,31,41). A case in point is gene regulation, since there are data indicating that Archaea use a transcription regulation machinery similar to that of eucaryotes (10,24,28,33). Whether this is the case also for archaeal heat shock genes has not been established. To address this question, it is necessary to first determine the expression pattern of the known archaeal heat shock genes and then proceed to analyze the regulation mechanisms at the molecular level.Among the Archaea, M. mazei is particularly interesting because it is one of a small group of organisms, the methanosarcinae (22,23), that undergo a growth cycle with morphologic conversions (3,25,32,42). The cycle includes a unicellular form or stage, named single cells, and two multicellular forms, called lamina and packets. The latter are globular structures with zonal heterogeneity and intercellular connective material that extends the outer surface of the packets and wraps the cells together. This connective material is composed chiefly of a heteropolysaccharide (14, 16) which confers to the packets resistance to traumas of various types: mechanical, physical (e.g., heat), and chemical (unpublished observations). This resistance is in contrast to the relative fragilit...

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confidence: 92%

Archaeal grpE: transcription in two different morphologic stages of Methanosarcina mazei and comparison with dnaK and dnaJ

1995

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“…Recently, part of the dnaK sequence was reported (26). We published its complete sequence (62) and that of the grpE gene (63). Here, we describe the isolation and the analysis of transcription of four heat shock genes which constitute the dnaK locus.…”

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confidence: 99%

Cloning, sequencing, and molecular analysis of the dnaK locus from Bacillus subtilis

et al. 1992

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By using an internal part of the dnaK gene from BaciUlus megaterium as a probe, a 5.2-kb Hindlll fragment of chromosomal DNA of BaciUlus subtilis was cloned. Downstream sequences were isolated by in vivo chromosome walking. Sequencing of 5,085 bp revealed four open reading frames in the order orf9-grpEdnaK-dnaJ. orJ39 encodes a 39-kDa polypeptide of unknown biological function with no noticeable homology to any other protein within the data bases. Alignment of the GrpE protein with those of three other bacterial species revealed a low overall homology, but a higher homology restricted to two regions which might be involved in interactions with other proteins. Alignment of the DnaK protein with six bacterial DnaK polypeptides revealed that a contiguous region of 24 amino acids is absent from the DnaK proteins of all known gram-positive species. Primer extension studies revealed three potential transcription start sites, two preceding orJ39 (Si and S2) and a third one in front of grpE (S3). S2 and S3 were activated at a high temperature. Northern (RNA) analysis led to the detection of three mRNA species of 4.9, 2.6, and 1.5 kb. RNA dot blot experiments revealed an at-least-fivefold increase in the amount of specific mRNA from 0 to 5 min postinduction and then a rapid decrease. A transcriptional fusion between dnaK and the amyL reporter gene exhibited a slight increase in a-amylase activity after heat induction. A 9-bp inverted repeat was detected in front of the coding region of orJ39. This inverted repeat is present in a number of other heat shock operons in other microorganisms ranging from cyanobacteria to mycobacteria. The biological property of this inverted repeat as a putative key element in the induction of heat shock genes is discussed. The dnaK locus was mapped at about 2230 on the B. subtilis genetic map.The heat shock response is a homeostatic mechanism that enables cells to survive a variety of environmental stresses. It is characterized by the increased synthesis of a group of evolutionarily conserved proteins, heat shock proteins (HSPs), and is a universal feature of both prokaryotic and eukaryotic cells (35). When Eschertichia coli cells are shifted to a high temperature, the synthesis of a set of about 20 HSPs transiently increases and then declines rapidly to steady-state levels characteristic of the new ambient temperature (44).The highly conserved HSPs perform similar functions in all organisms. One of these functions is the well-established regulation of protein-protein interactions by the chaperonins (69). One of the most abundant HSPs, HSP70, is highly conserved in evolution. It is found in such diverse organisms as E. coli, Saccharomyces cerevisiae, Drosophila melanogaster, and Homo sapiens (14, 35).The dnaK gene of E. coli was originally discovered because mutations in it blocked bacteriophage lambda DNA replication at all temperatures (22,23). Subsequently, the dnaK gene product was shown to be essential for E. coli viability at high and low temperatures (22,30,46,51,52), and genetic evid...

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“…Class I genes are specifically induced by heat stress (17). The well-known chaperonins GroEL, GroES, DnaK, DnaJ, and GrpE are encoded by genes belonging to this group (30,41,48,54,55). The transcription of the respective genes is regulated by HrcA, a transcription repressor which binds to the CIRCE element (43,56,57,59).…”

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Transcription of the nfrA-ywcH Operon from Bacillus subtilis Is Specifically Induced in Response to Heat

2000

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The NfrA protein, an oxidoreductase from the soil bacterium Bacillus subtilis, is synthesized during the stationary phase and in response to heat. Analysis of promoter mutants revealed that the nfrA gene belongs to the class III heat shock genes in B. subtilis. An approximate 10-fold induction at both the transcriptional and the translational levels was found after thermal upshock. This induction resulted from enhanced synthesis of mRNA. Genetic and Northern blot analyses revealed that nfrA and the gene downstream of nfrA are transcribed as a bicistronic transcriptional unit. The unstable full-length transcript is processed into two short transcripts encoding nfrA and ywcH. The nfrA-ywcH operon is not induced by salt stress or by ethanol. According to previously published data, the transcription of class III genes in general is activated in response to the addition of these stressors. However, this conclusion is based on experiments which lacked a valid control. Therefore, it seems possible that the transcription of all class III genes is specifically induced by heat shock.The continuous demand of Bacillus subtilis to adapt to everchanging conditions in its natural environment has forced the generation of complex regulatory mechanisms governing the transcription of stress-specific proteins. Stress-inducible genes from B. subtilis in general are subdivided into three groups (17,18). Class I genes are specifically induced by heat stress (17). The well-known chaperonins GroEL, GroES, DnaK, DnaJ, and GrpE are encoded by genes belonging to this group (30,41,48,54,55). The transcription of the respective genes is regulated by HrcA, a transcription repressor which binds to the CIRCE element (43,56,57,59). Genes transcribed in a Bdependent manner constitute class II stress-responsive genes (17, 18). B activity is triggered by different kinds of stress and by starvation (5, 7-9). Members of the last group of stressinduced genes, class III, are induced not by starvation but by several different stressful conditions. The transcription of class III genes is neither repressed by HrcA nor solely dependent on B . The regulator of the clpC operon, which encodes class III proteins, is known (11,27). This operon is transcribed by the activity of RNA polymerases containing B and A (28). Nevertheless, transcription is not induced at the onset of the stationary phase (28), likely because of the activity of this regulator (11,27).Some of the stress-responsive proteins are regulated by two transcription factors. clpC, dps, trxA, opuE, and clpP (1,4,15,29,40,46) are transcribed by RNA polymerase containing either A or B . csbB is under the additional control of X (22). The csb40 operon (50) and the yvyD gene (13) are transcribed from B and H promoters, respectively. This genetic organization enables the bacterial cell to modulate the regulation of the respective genes in response to additional challenges.In this communication, we describe the transcriptional regulation of the nfrA-ywcH operon encoding an oxidoreductase (34, 58) and a p...

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Nucleotide sequence of aBacillus subtilisgene homologous to thegrpEgene ofE. colilocated immediately upstream of thednaKgene

Cited by 25 publications

References 4 publications

Archaeal grpE: transcription in two different morphologic stages of Methanosarcina mazei and comparison with dnaK and dnaJ

Archaeal grpE: transcription in two different morphologic stages of Methanosarcina mazei and comparison with dnaK and dnaJ

Cloning, sequencing, and molecular analysis of the dnaK locus from Bacillus subtilis

Transcription of the nfrA-ywcH Operon from Bacillus subtilis Is Specifically Induced in Response to Heat

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