The universal stress protein A of Escherichia coli is required for resistance to DNA damaging agents and is regulated by a RecA/FtsK‐dependent regulatory pathway

The link between cell division defects and the induction of the universal stress response is demonstrated to operate via the RecA regulator of the SOS response. An insertion in the cell division gene ftsK upregulates uspA in a recA‐dependent manner. Unlike true SOS response genes, this upregulation only occurs in growth‐arrested cells and is LexA independent. Thus, besides ppGpp‐dependent starvation signals, DNA aberrations transduce RecA‐dependent signals to the uspA promoter, which only affect the promoter during stasis. Further, we show that ftsK itself, like uspA, is induced in stationary phase and that this induction requires the stringent control modulon rather than activated RecA. Thus, ftsK, like uspA, is regulated by at least two global regulators: ppGpp of the stringent control network and RecA of the SOS modulon. We suggest that UspA is a new bona fide member of the RecA‐dependent DNA protection and repair system, as mutants lacking functional UspA were found to be sensitive to UV irradiation and mitomycin C exposure. Moreover, the UV sensitivity of uspA mutants is enhanced in an additive manner by the ftsK1 mutation.

[1]  C. Thompson,et al.  Characterization of dinY, a new Escherichia coli DNA repair gene whose products are damage inducible even in a lexA(Def) background , 1993, Journal of bacteriology.

[2]  R. D'ari,et al.  Analysis of the effect of ppGpp on the ftsQAZ operon in Escherichia coli , 1998, Molecular microbiology.

[3]  T. Nyström,et al.  Expression and role of the universal stress protein, UspA, of Escherichia coli during growth arrest , 1994, Molecular microbiology.

[4]  J. Errington,et al.  Bacillus subtilis SpoIIIE protein required for DNA segregation during asymmetric cell division. , 1994, Science.

[5]  T. Nyström,et al.  Effects of starvation for exogenous carbon on functional mRNA stability and rate of peptide chain elongation in Escherichia coli. , 1994, FEMS microbiology letters.

[6]  Mutations at Pro67in the RecA Protein P-loop Motif Differentially Modify Coprotease Function and Separate Coprotease from Recombination Activities(*) , 1995, The Journal of Biological Chemistry.

[7]  P. O’Farrell.  High resolution two-dimensional electrophoresis of proteins. , 1975, The Journal of biological chemistry.

[8]  K.,et al.  Isolation of DNA damage-inducible promoters in Escherichia coli: regulation of polB (dinA), dinG, and dinH by LexA repressor , 1992, Journal of bacteriology.

[9]  T. Nyström.  The trials and tribulations of growth arrest. , 1995, Trends in microbiology.

[10]  W. Donachie,et al.  A new Escherichia coli cell division gene, ftsK , 1995, Journal of bacteriology.

[11]  F. Neidhardt,et al.  Ribosomes as sensors of heat and cold shock in Escherichia coli. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[12]  J. Errington,et al.  Postseptational chromosome partitioning in bacteria. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[13]  R. Kolter,et al.  DNA protection by stress-induced biocrystallization , 1999, Nature.

[14]  W. Donachie,et al.  Only the N-Terminal Domain of FtsK Functions in Cell Division , 1998, Journal of bacteriology.

[15]  G. Ruhnke,et al.  The effects of starvation. , 1949 .

[16]  T. Nyström,et al.  Cloning, mapping and nucleotide sequencing of a gene encoding a universal stress protein in Eschericha coli , 1992, Molecular microbiology.

[17]  P. Freestone,et al.  Protein phosphorylation in Escherichia coli L. form NC-7. , 1998, Microbiology.

[18]  R. D'ari,et al.  Effect of suppressors of SOS-mediated filamentation on sfiA operon expression in Escherichia coli , 1983, Journal of bacteriology.

[19]  W. Donachie,et al.  FtsK is a bifunctional protein involved in cell division and chromosome localization in Escherichia coli , 1998, Molecular microbiology.

[20]  T. Nyström,et al.  The universal stress protein, UspA, of Escherichia coli is phosphorylated in response to stasis. , 1997, Journal of molecular biology.

[21]  D. Jin,et al.  The rpoB mutants destabilizing initiation complexes at stringently controlled promoters behave like "stringent" RNA polymerases in Escherichia coli. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[22]  D. Mount,et al.  Cleavage of the Escherichia coli lexA protein by the recA protease. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[23]  J. Lutkenhaus,et al.  FtsK is an essential cell division protein that is localized to the septum and induced as part of the SOS response , 1998, Molecular microbiology.

[24]  M. Aldea,et al.  Division genes in Escherichia coli are expressed coordinately to cell septum requirements by gearbox promoters. , 1990, The EMBO journal.

[25]  W. Donachie,et al.  The cytoplasmic domain of FtsK protein is required for resolution of chromosome dimers , 1999, Molecular microbiology.

[26]  R. Kolter,et al.  An E. coli promoter induced by the cessation of growth , 1987, Molecular microbiology.

[27]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[28]  T. Linn,et al.  Improved vector system for constructing transcriptional fusions that ensures independent translation of lacZ , 1990, Journal of bacteriology.

[29]  T. Nyström,et al.  Tyrosine phosphorylation in Escherichia coli. , 1998, Journal of molecular biology.

[30]  E V Koonin,et al.  Sequence analysis of eukaryotic developmental proteins: ancient and novel domains. , 1996, Genetics.

[31]  T. Nyström,et al.  Effects of overproducing the universal stress protein, UspA, in Escherichia coli K-12 , 1996, Journal of bacteriology.

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

[33]  T. Nyström,et al.  Emergency derepression: stringency allows RNA polymerase to override negative control by an active repressor , 2000, Molecular microbiology.

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

[35]  W. Margolin,et al.  Role of the C terminus of FtsK in Escherichia coli chromosome segregation. , 1998, Journal of bacteriology.

[36]  T Nyström,et al.  Maintenance energy requirement: what is required for stasis survival of Escherichia coli? , 1998, Biochimica et biophysica acta.

[37]  T. Nyström,et al.  A mutation in the ftsK gene of Escherichia coli affects cell-cell separation, stationary-phase survival, stress adaptation, and expression of the gene encoding the stress protein UspA , 1997, Journal of bacteriology.

[38]  C. Cayrol,et al.  Recovery of respiration following the SOS response of Escherichia coli requires RecA-mediated induction of 2-keto-4-hydroxyglutarate aldolase. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[39]  D. Sherratt,et al.  FtsK‐dependent and ‐independent pathways of Xer site‐specific recombination , 1999, The EMBO journal.

[40]  T. Nyström,et al.  Isolation and properties of a mutant of Escherichia coli with an insertional inactivation of the uspA gene, which encodes a universal stress protein , 1993, Journal of bacteriology.

[41]  T. Nyström,et al.  Role of the Escherichia coli FadR regulator in stasis survival and growth phase-dependent expression of the uspA, fad, and fab genes , 1996, Journal of bacteriology.