Mechanism of CRP-cAMP activation of lac operon transcription initiation activation of the P1 promoter.

CRP-cAMP was shown to activate transcription initiation at the Escherichia coli lac promoter in vitro as a result of two separate effects. An indirect component of the activation resulted from an enhancement of the fraction of promoters productively bound by RNA polymerase. This effect was due largely to CRP-cAMP repression of RNA polymerase binding to an overlapping site (lac P2) within the promoter region. In addition, a direct enhancement of RNA polymerase binding at the principal lac promoter (lac P1) was found. The combination of indirect and direct activation by CRP-cAMP was suggested to be responsible for the large activation observed in vivo. Promoter strength parameters were also determined for the L8, UV5 and Ps promoters. The effect of CRP-cAMP on these mutant promoters was shown to be consistent with the activation mechanism deduced for the lac wild-type promoter. DNA supercoiling enhanced the promoter strength of the lac wild-type and UV5 promoters. The combination of supercoiling and CRP-cAMP was necessary for optimal promoter strength for the lac wild-type promoter.

[1]  W. Reznikoff,et al.  Genetic regulation: the Lac control region. , 1975, Science.

[2]  W. McClure,et al.  Studies on the selectivity of DNA precipitation by spermine. , 1981, Nucleic Acids Research.

[3]  M. Chamberlin,et al.  A quantitative assay for bacterial RNA polymerases. , 1979, The Journal of biological chemistry.

[4]  J. Lis.  Fractionation of DNA fragments by polyethylene glycol induced precipitation. , 1980, Methods in enzymology.

[5]  Diane K. Hawley,et al.  The effect of a lambda repressor mutation on the activation of transcription initiation from the lambda P RM promoter , 1983, Cell.

[6]  J. Beckwith,et al.  Evidence for two sites in the lac promoter region. , 1972, Journal of molecular biology.

[7]  H. Buc,et al.  Comparison of the binding sites for the Escherichia coli cAMP receptor protein at the lactose and galactose promoters. , 1983, The EMBO journal.

[8]  M G Fried,et al.  CAP and RNA polymerase interactions with the lac promoter: binding stoichiometry and long range effects , 1983, Nucleic Acids Res..

[9]  R. Sinden,et al.  Torsional tension in the DNA double helix measured with trimethylpsoralen in living E. coli cells: Analogous measurements in insect and human cells , 1980, Cell.

[10]  W. Gilbert,et al.  Sequencing end-labeled DNA with base-specific chemical cleavages. , 1980, Methods in enzymology.

[11]  H. Buc,et al.  Is DNA unwound by the cyclic AMP receptor protein? , 1982, Nucleic acids research.

[12]  W R McClure,et al.  Characterization of ribonucleic acid polymerase-T7 promoter binary complexes. , 1980, Biochemistry.

[13]  M J Chamberlin,et al.  The selectivity of transcription. , 1974, Annual review of biochemistry.

[14]  J D Gralla,et al.  Mutation-induced changes in RNA polymerase-lac ps promoter interactions. , 1982, The Journal of biological chemistry.

[15]  D. E. Johnston,et al.  A steady state assay for the RNA polymerase initiation reaction. , 1978, The Journal of biological chemistry.

[16]  A. Schmitz.  Cyclic AMP receptor proteins interacts with lactose operator DNA. , 1981, Nucleic acids research.

[17]  R. Burgess,et al.  Purification and properties of the sigma subunit of Escherichia coli DNA-dependent RNA polymerase. , 1979, Biochemistry.

[18]  L. Maquat,et al.  In vitro analysis of the Escherichia coli RNA polymerase interaction with wild-type and mutant lactose promoters. , 1978, Journal of molecular biology.

[19]  A. Revzin,et al.  Stoichiometry of catabolite activator protein/adenosine cyclic 3',5'-monophosphate interactions at the lac promoter of Escherichia coli. , 1982, Biochemistry.

[20]  William R. McClure,et al.  Dual promoter control of the escherichia coli lactose operon , 1984, Cell.

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

[22]  Keith R. Yamamoto,et al.  Biological Regulation and Development , 1982, Springer US.

[23]  R. Burgess,et al.  A procedure for the rapid, large-scall purification of Escherichia coli DNA-dependent RNA polymerase involving Polymin P precipitation and DNA-cellulose chromatography. , 1975, Biochemistry.

[24]  H. Buc,et al.  Point mutations change the thermal denaturation profile of a short DNA fragment containing the lactose control elements. Comparison between experiment and theory. , 1982, The EMBO journal.

[25]  Thomas A. Steitz,et al.  Structure of catabolite gene activator protein at 2.9 Å resolution suggests binding to left-handed B-DNA , 1981, Nature.

[26]  M. Rosenberg,et al.  A promoter of pBR322 activated by cAMP receptor protein. , 1981, Nucleic acids research.

[27]  H. Riezman,et al.  Transcription and translation initiation frequencies of the Escherichia coli lac operon. , 1977, Journal of molecular biology.

[28]  B. Sanzey,et al.  Modulation of gene expression by drugs affecting deoxyribonucleic acid gyrase , 1979, Journal of bacteriology.

[29]  T. Steitz,et al.  Structure of catabolite gene activator protein at 2.9-A resolution. Incorporation of amino acid sequence and interactions with cyclic AMP. , 1982, The Journal of biological chemistry.

[30]  N. Davidson.  Effect of DNA length on the free energy of binding of an unwinding ligand to a supercoiled DNA. , 1972, Journal of molecular biology.

[31]  J. Majors,et al.  Specific binding of CAP factor to lac promoter DNA , 1975, Nature.