The rpoE gene of Escherichia coli, which encodes sigma E, is essential for bacterial growth at high temperature

In vitro transcription analysis has shown that only RNA polymerase containing an alternative sigma subunit, sigma E, activates transcription from one of the rpoH promoters and the htrA promoter. The location of the rpoE gene encoding sigma E on the Escherichia coli chromosome has recently been established, but no rpoE mutant has yet become available for phenotypic testing. We cloned the rpoE gene from the lambda-ordered clones of the E. coli genome and confirmed that the reconstituted RNA polymerase containing the gene product (E sigma E) can transcribe htrA in vitro. We constructed an rpoE-defective strain by gene disruption using the cloned rpoE gene. We demonstrate that expression of htrA is completely dependent on the rpoE gene in vivo and that the rpoE gene is essential for bacterial growth at high temperature.

[1]  Jeffrey H. Miller.  Experiments in molecular genetics , 1972 .

[2]  E. Southern.  Detection of specific sequences among DNA fragments separated by gel electrophoresis. , 1975, Journal of molecular biology.

[3]  J. Vieira,et al.  The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. , 1982, Gene.

[4]  K. Ito,et al.  Heat shock regulatory gene (htpR) of Escherichia coli is required for growth at high temperature but is dispensable at low temperature. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[5]  A. Grossman,et al.  The htpR gene product of E. coli is a sigma factor for heat-shock promoters , 1984, Cell.

[6]  E. Craig,et al.  The heat shock response. , 1985, CRC critical reviews in biochemistry.

[7]  S. Elledge,et al.  Site-directed insertion and deletion mutagenesis with cloned fragments in Escherichia coli , 1985, Journal of bacteriology.

[8]  F. Studier,et al.  Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. , 1986, Journal of molecular biology.

[9]  F. Neidhardt,et al.  Escherichia Coli and Salmonella: Typhimurium Cellular and Molecular Biology , 1987 .

[10]  Carol A. Gross,et al.  The heat shock response of E. coli is regulated by changes in the concentration of σ32 , 1987, Nature.

[11]  F. Neidhardt,et al.  Regulation of the promoters and transcripts of rpoH, the Escherichia coli heat shock regulatory gene. , 1987, Genes & development.

[12]  K. Isono,et al.  The physical map of the whole E. coli chromosome: Application of a new strategy for rapid analysis and sorting of a large genomic library , 1987, Cell.

[13]  R. Simons,et al.  Improved single and multicopy lac-based cloning vectors for protein and operon fusions. , 1987, Gene.

[14]  C. Gross,et al.  Escherichia coli heat shock gene mutants are defective in proteolysis. , 1988, Genes & development.

[15]  C. Georgopoulos,et al.  Sequence analysis and regulation of the htrA gene of Escherichia coli: a sigma 32-independent mechanism of heat-inducible transcription. , 1988, Nucleic acids research.

[16]  M. Chamberlin,et al.  Structure and function of bacterial sigma factors. , 1988, Annual review of biochemistry.

[17]  J. Kaguni,et al.  A novel sigma factor is involved in expression of the rpoH gene of Escherichia coli , 1989, Journal of bacteriology.

[18]  C. Gross,et al.  Identification of the sigma E subunit of Escherichia coli RNA polymerase: a second alternate sigma factor involved in high-temperature gene expression. , 1989, Genes & development.

[19]  C. Georgopoulos,et al.  Identification, characterization, and mapping of the Escherichia coli htrA gene, whose product is essential for bacterial growth only at elevated temperatures , 1989, Journal of bacteriology.

[20]  D. Shoemaker,et al.  High-fidelity amplification using a thermostable DNA polymerase isolated from Pyrococcus furiosus. , 1991, Gene.

[21]  A. Ishihama,et al.  Bipartite functional map of the E. coli RNA polymerase α subunit: Involvement of the C-terminal region in transcription activation by cAMP-CRP , 1991, Cell.

[22]  F. Hartl,et al.  Prevention of protein denaturation under heat stress by the chaperonin Hsp60. , 1992, Science.

[23]  H. Echols,et al.  Activity of the Hsp70 chaperone complex--DnaK, DnaJ, and GrpE--in initiating phage lambda DNA replication by sequestering and releasing lambda P protein. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[24]  DnaK and DnaJ heat shock proteins participate in protein export in Escherichia coli. , 1992, Genes & development.

[25]  H. Mori,et al.  Regulation of the heat-shock response in bacteria. , 1993, Annual review of microbiology.

[26]  K. Makino,et al.  Role of the sigma 70 subunit of RNA polymerase in transcriptional activation by activator protein PhoB in Escherichia coli. , 1993, Genes & development.

[27]  D. Martin,et al.  Conversion of Pseudomonas aeruginosa to mucoidy in cystic fibrosis: environmental stress and regulation of bacterial virulence by alternative sigma factors , 1994, Journal of bacteriology.

[28]  K. Rudd,et al.  Analysis of the Streptomyces coelicolor sigE gene reveals the existence of a subfamily of eubacterial RNA polymerase sigma factors involved in the regulation of extracytoplasmic functions. , 1994, Proceedings of the National Academy of Sciences of the United States of America.