Stimulation of the phage λ pL promoter by integration host factor requires the carboxy terminus of the α-subunit of RNA polymerase

Abstract Escherichia coli integration host factor (IHF) binds with high affinity to two tandem IHF consensus sequences located upstream from the pL promoter of bacteriophage lambda. IHF was shown to stimulate transcription initiation from the pL promoter by increasing close complex formation (KB). We show here, by the use of reconstituted mutant RNA polymerases, that the C-terminal portion of the α subunit of RNA polymerase plays an essential role in the stimulation of transcription by IHF. Our results are in agreement with the hypothesis that IHF, like the cAMP-CRP activator, increases the affinity of RNA polymerase to the promoter by protein-protein interaction.

[1]  A. Ishihama,et al.  Functional specialization within the α-subunit of Escherichia coli RNA polymerase , 1991 .

[2]  R. Weisberg,et al.  Overproduction of Escherichia coli integration host factor, a protein with nonidentical subunits , 1987, Journal of bacteriology.

[3]  S. Adhya,et al.  Cooperative DNA binding of heterologous proteins: evidence for contact between the cyclic AMP receptor protein and RNA polymerase. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[4]  S. Kustu,et al.  The integration host factor stimulates interaction of RNA polymerase with NIFA, the transcriptional activator for nitrogen fixation operons , 1990, Cell.

[5]  L. Shapiro,et al.  Integration host factor is required for the activation of developmentally regulated genes in Caulobacter. , 1990, Genes & development.

[6]  Diane K. Hawley,et al.  Mechanism of activation of transcription initiation from the λPRM promoter , 1982 .

[7]  A. Oppenheim,et al.  Supercoiling, integration host factor, and a dual promoter system, participate in the control of the bacteriophage λ pL promoter☆ , 1992 .

[8]  H. Buc,et al.  On the action of the cyclic AMP‐cyclic AMP receptor protein complex at the Escherichia coli lactose and galactose promoter regions. , 1984, The EMBO journal.

[9]  H. Krause,et al.  Positive and negative regulation of the Mu operator by Mu repressor and Escherichia coli integration host factor. , 1986, The Journal of biological chemistry.

[10]  A. Ishihama,et al.  Subunit of assembly of Escherichia coli RNA polymerase. , 1981, Advances in biophysics.

[11]  K. Makino,et al.  Functional map of the alpha subunit of Escherichia coli RNA polymerase: two modes of transcription activation by positive factors. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[12]  P. van de Putte,et al.  Integration host factor of Escherichia coli regulates early- and repressor transcription of bacteriophage Mu by two different mechanisms. , 1988, Nucleic acids research.

[13]  T. Silhavy,et al.  Suppressor mutations in rpoA suggest that OmpR controls transcription by direct interaction with the alpha subunit of RNA polymerase , 1991, Journal of bacteriology.

[14]  P. van de Putte,et al.  Analysis of the IHF binding site in the regulatory region of bacteriophage Mu. , 1991, Nucleic acids research.

[15]  D M Crothers,et al.  Synergy between Escherichia coli CAP protein and RNA polymerase in the lac promoter open complex. , 1989, Journal of molecular biology.

[16]  H. Nakamura,et al.  In vivo enhancement of general and specific transcription in Escherichia coli by DNA gyrase activity. , 1979, Gene.

[17]  M. Ptashne,et al.  Repressor structure and the mechanism of positive control , 1983, Cell.

[18]  W. Reznikoff,et al.  Mutations in the lac P2 promoter , 1987, Journal of bacteriology.

[19]  D. Friedman,et al.  Integration host factor: A protein for all reasons , 1988, Cell.

[20]  B. Magasanik,et al.  Role of integration host factor in the regulation of the glnHp2 promoter of Escherichia coli. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[21]  C. Miller,et al.  Mutations in rpoA affect expression of anaerobically regulated genes in Salmonella typhimurium , 1991, Journal of bacteriology.

[22]  G W Hatfield,et al.  Characterization of the integration host factor binding site in the ilvPG1 promoter region of the ilvGMEDA operon of Escherichia coli. , 1990, The Journal of biological chemistry.

[23]  M. Freundlich,et al.  Integration host factor is a negative effector of in vivo and in vitro expression of ompC in Escherichia coli , 1990, Journal of bacteriology.

[24]  Kevin Gaston,et al.  Stringent spacing requirements for transcription activation by CRP , 1990, Cell.

[25]  K. Timmis,et al.  An upstream XylR‐ and IHF‐induced nucleoprotein complex regulates the sigma 54‐dependent Pu promoter of TOL plasmid. , 1991, The EMBO journal.

[26]  H. Buc,et al.  Overlapping promoters and their control in Escherichia coli: the gal case. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[27]  J. Kincade,et al.  Bacteriophage lambda promoters pL and pR: sequence determinants of in vivo activity and of sensitivity to the DNA gyrase inhibitor, coumermycin. , 1991, Gene.

[28]  A. Oppenheim,et al.  Integration host factor stimulates the phage lambda pL promoter. , 1990, Journal of molecular biology.

[29]  A. Oppenheim,et al.  Repression of the λ pcin promoter by integrative host factor , 1989 .

[30]  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.

[31]  W. McClure,et al.  Mechanism of CRP-cAMP activation of lac operon transcription initiation activation of the P1 promoter. , 1984, Journal of molecular biology.

[32]  R. F. Pereira,et al.  Binding of integration host factor (IHF) to the ilvGp1 promoter of the ilvGMEDA operon of Escherichia coli K12 , 1988, Nucleic Acids Res..

[33]  N. Fujita,et al.  Sequence analysis of two temperature-sensitive mutations in the alpha subunit gene (rpoA) of Escherichia coli RNA polymerase. , 1990, Nucleic acids research.

[34]  M. Freundlich,et al.  Altered osmoregulation of ompF in integration host factor mutants of Escherichia coli , 1988, Journal of bacteriology.

[35]  Integration host factor-mediated expression of the ilvGMEDA operon of Escherichia coli. , 1991, The Journal of biological chemistry.

[36]  W. Szybalski,et al.  Physical and biological consequences of interactions between integration host factor (IHF) and coliphage lambda late p'R promoter and its mutants. , 1989, Gene.

[37]  J. Tomizawa.  Control of ColE1 plasmid replication: Binding of RNA I to RNA II and inhibition of primer formation , 1986, Cell.