In vitro effect of the Escherichia coli heat shock regulatory protein on expression of heat shock genes

Bloom, Mark; Skelly, Susan; VanBogelen, Ruth A.; Neidhardt, Frederick C.; Brot, Nathan; Weissbach, Herbert

doi:10.1128/jb.166.2.380-384.1986

Cited by 49 publications

(34 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…ArpoH (formerly referred to as htpR) strains, which are not viable at temperatures of >20°C, have decreased expression of GroES/EL and probably contain little or no DnaK (4,26,60). Expression of DnaK from the tryptophan promoter alone did not substantially increase the upper growth temperature of these ArpoH cells.…”

Section: Discussionmentioning

confidence: 99%

DnaK mutants defective in ATPase activity are defective in negative regulation of the heat shock response: expression of mutant DnaK proteins results in filamentation

McCarty

Walker

1994

J Bacteriol

View full text Add to dashboard Cite

Site-directed mutagenesis has previously been used to construct Escherichia coli dnaK mutants encoding proteins that are altered at the site of in vitro phosphorylation (J. S. McCarty and G. C. Walker, Proc. Natl. Acad. Sci. USA 88:9513-9517, 1991 Escherichia coli, in common with many organisms, responds to sudden shifts in temperature by transiently inducing members of the heat shock regulon, which consists of genes and operons that are under the transcriptional regulation of the heat shock sigma factor 32 (42). By genetic analysis, this regulon has been shown to be negatively regulated by three of these heat shock proteins, DnaK, DnaJ, and GrpE. dnaK, dnaJ, and grpE mutants have the phenotype of overproducing the proteins of the heat shock regulon (50). The primary mechanisms for this negative regulation appear to be effects on the level of synthesis, stability, and activity of c32 (19,(49)(50)(51)53

show abstract

Section: Discussionmentioning

confidence: 99%

DnaK mutants defective in ATPase activity are defective in negative regulation of the heat shock response: expression of mutant DnaK proteins results in filamentation

McCarty

Walker

1994

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…These genes are dispersed throughout the chromosome, and their products perform various functions in the cell, most of which are not clearly understood thus far (12,16,30,31,34). The transcription of the majority of these genes is positively regulated by the product of the rpoH gene (previously known also as htpR or hin; for a review, see references 30 and 31), the ar32 subunit of the RNA polymerase holoenzyme (6,17). Some of these genes are also essential for bacterial growth under normal temperature conditions, for example, the rpoD gene, which codes for the cr70 subunit of RNA polymerase (30,31), or grpE, which is essential for the replication of bacteriophage X but which plays an otherwise unknown role in E. coli physiology (D. Ang and C. Georgopoulos, submitted for publication).…”

mentioning

confidence: 99%

Identification, characterization, and mapping of the Escherichia coli htrA gene, whose product is essential for bacterial growth only at elevated temperatures

et al. 1989

View full text Add to dashboard Cite

As a typical mesophile, Escherichia coli can maintain balanced growth between approximately 10 and 49°C (19). In the range of approximately 21 to 37°C, the rate of E. coli growth varies as a simple function of temperature (19). Raising the temperature above 40°C leads to progressively slower growth rates and changes in the cellular content of many proteins (30, 31). The adaptation processes that occur on a shift to high temperature include an increased expression of a set of genes, called heat shock genes, many of which are highly conserved among procaryotic and eucaryotic organisms. These genes are dispersed throughout the chromosome, and their products perform various functions in the cell, most of which are not clearly understood thus far (12,16,30,31,34). The transcription of the majority of these genes is positively regulated by the product of the rpoH gene (previously known also as htpR or hin; for a review, see references 30 and 31), the ar32 subunit of the RNA polymerase holoenzyme (6,17). Some of these genes are also essential for bacterial growth under normal temperature conditions, for example, the rpoD gene, which codes for the cr70 subunit of RNA polymerase (30, 31), or grpE, which is essential for the replication of bacteriophage X but which plays an otherwise unknown role in E. coli physiology (D. Ang and C. Georgopoulos, submitted for publication). Several heat shock genes, like lysU, which codes for an alternate form of lysyl-tRNA synthetase (30,31,38), or lon, which codes for an ATP-dependent protease (26), are not absolutely essential for bacterial growth. There is also a class of heat shock genes that is conditionally dispensable at low temperatures; i.e., deletion mutants can be constructed at low temperatures but they grow poorly and rapidly accumulate extragenic suppressors (e.g., dnaK [7a] and dnaJ [S. Sell and C. Georgopoulos, unpublished data]).Apart from the canonical heat shock genes, the rpoH regulatory gene itself is indispensable for cell adaptation to high temperatures (9). It is also known from two-dimensional electrophoresis of total E. coli proteins that there are other * Corresponding author.

show abstract

“…Four promoters, designated rpoHip, rpoH2p, rpoH3p, and rpoH4p, have been mapped at about 220, 130, 90, and 80 base pairs, respectively, upstream from the coding sequence (10,11). Promoters rpoHlp and rpoH4p are recognized by U70 RNA polymerase (Eu70) (3,10,11). A third promoter, rpoH2p, appears to be strain specific.…”

mentioning

confidence: 99%

A novel sigma factor is involved in expression of the rpoH gene of Escherichia coli

1989

View full text Add to dashboard Cite

The Escherichia coli rpoH gene encoding J32, which is involved in the heat shock response, is transcribed from as many as four promoters. We have isolated a novel sigma factor of about 24 kilodaltons that allows core RNA polymerase to transcribe preferentially from one of these promoters, rpoH3p. This promoter is known to be regulated by DnaA protein. The,24 factor was isolated from a preparation of RNA polymerase by electroelution from sodium dodecyl sulfate-polyacrylamide gels followed by renaturation. Expression of heat shock proteins is induced by treatments which include those that induce the stringent response. Under such conditions, decreased transcription from rpoH3p and no increase in transcription from other rpoH promoters were observed. This result suggests that induction of heat shock proteins by the stringent response is not mediated by increased transcription of the rpoH gene. (8,14,21). Upon temperature upshift, the increased expression of heat shock proteins is attributable to an increase in u32 due to increased expression and stability (10,11,31,32). A reprogramming of RNA polymerase ensues.Receht experiments have examined transcription of the rpoH gene (10,11,32). Four promoters, designated rpoHip, rpoH2p, rpoH3p, and rpoH4p, have been mapped at about 220, 130, 90, and 80 base pairs, respectively, upstream from the coding sequence (10, 11). Promoters rpoHlp and rpoH4p are recognized by U70 RNA polymerase (Eu70) (3, 10, 11). A third promoter, rpoH2p, appears to be strain specific. In vitro transcription from the fourth promoter, rpoH3p, does not involve Eu70 or Eu32 (3, 10). These results suggest that a novel sigma factor or a positive regulatory protein may be required for transcription from rpoH3p. In related experiments examining regulation of the rpoH gene, we identified a preparation of RNA polymerase able to initiate transcription from rpoHip, rpoH3p, and rpoH4p (33a). From this, we have isolated a protein of 24 kilodaltons (kDa) which confers rpoH3p-specific transcription to core RNA polymerase. We also report on transcriptional regulation of the rpoH gene during the stringent response. MATERIALS AND METHODSBacterial strains and plasmids. E. coli W3110 and AB1157 thr-J ara-14 leuB6 (gpt-proA)62 lacYl tsx-33 supE44 galK2 hisG4 rpsL31 xyl-5 mtl-l argE3 thi-J were from the E. coli Genetic Stock Center. Plasmid pFN82 was from Frederick C. Neidhardt (University of Michigan). To construct this plasmid, a 2.5-kilobase-pair PstI fragment containing the rpoH promoter region and adjacent N-terminal coding region was inserted at the single PstI site of pBR322 (27)

show abstract

In vitro effect of the Escherichia coli heat shock regulatory protein on expression of heat shock genes

Cited by 49 publications

References 30 publications

DnaK mutants defective in ATPase activity are defective in negative regulation of the heat shock response: expression of mutant DnaK proteins results in filamentation

DnaK mutants defective in ATPase activity are defective in negative regulation of the heat shock response: expression of mutant DnaK proteins results in filamentation

Identification, characterization, and mapping of the Escherichia coli htrA gene, whose product is essential for bacterial growth only at elevated temperatures

A novel sigma factor is involved in expression of the rpoH gene of Escherichia coli

Contact Info

Product

Resources

About