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. 1982 Jun 25;10(12):3791–3802. doi: 10.1093/nar/10.12.3791

Structure of the complex between lac repressor headpiece and operator DNA from measurements of the orientation relaxation and the electric dichroism.

D Pörschke, N Geisler, W Hillen
PMCID: PMC320753  PMID: 7050912

Abstract

The complex between lac repressor headpiece and short rodlike DNA fragments containing the lac operator sequence is characterised by measurements of the rotation diffusion. Using the method of electric dichroism we measure the rotation relaxation and determine changes in the length of the DNA upon ligand binding with high accuracy. According to these measurements any change in the length of the operator DNA upon binding of the first two headpiece molecules remains below 1A; the electric dichroism also remains virtually unchanged. At high degrees of (unspecific) binding we observe an increase in the rotation relaxation time, which is attributed to an increase of the apparent mean radius of the complex. As a control of our procedure for the determination of length changes we use the intercalation of ethidium bromide and arrive at an increase of the DNA length per bound ethidium of 3.2A (at 3.4A rise per base pair). The results obtained for the headpiece operator complex are not consistent with models assuming large changes of the DNA structure or intercalation of tyrosine residues.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bourgeois S., Pfahl M. Repressors. Adv Protein Chem. 1976;30:1–99. doi: 10.1016/s0065-3233(08)60478-7. [DOI] [PubMed] [Google Scholar]
  2. Charlier M., Maurizot J. C., Zaccai G. Neutron-scattering studies of lac repressor: a low-resolution model. J Mol Biol. 1981 Nov 25;153(1):177–182. doi: 10.1016/0022-2836(81)90533-7. [DOI] [PubMed] [Google Scholar]
  3. Dunaway M., Manly S. P., Matthews K. S. Model for lactose repressor protein and its interaction with ligands. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7181–7185. doi: 10.1073/pnas.77.12.7181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Geisler N., Weber K. Isolation of a set of hybrid lac repressors made in vitro between normal lac repressor and its homogeneous tryptic core. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3103–3106. doi: 10.1073/pnas.73.9.3103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Geisler N., Weber K. Isolation of amino-terminal fragment of lactose repressor necessary for DNA binding. Biochemistry. 1977 Mar 8;16(5):938–943. doi: 10.1021/bi00624a020. [DOI] [PubMed] [Google Scholar]
  6. Gilbert W., Maxam A. The nucleotide sequence of the lac operator. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3581–3584. doi: 10.1073/pnas.70.12.3581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hagerman P. J. Investigation of the flexibility of DNA using transient electric birefringence. Biopolymers. 1981 Jul;20(7):1503–1535. doi: 10.1002/bip.1981.360200710. [DOI] [PubMed] [Google Scholar]
  8. Hillen W., Klein R. D., Wells R. D. Preparation of milligram amounts of 21 deoxyribonucleic acid restriction fragments. Biochemistry. 1981 Jun 23;20(13):3748–3756. doi: 10.1021/bi00516a013. [DOI] [PubMed] [Google Scholar]
  9. Hogan M., Dattagupta N., Crothers D. M. Transient electric dichroism of rod-like DNA molecules. Proc Natl Acad Sci U S A. 1978 Jan;75(1):195–199. doi: 10.1073/pnas.75.1.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hogan M., Dattagupta N., Crothers D. M. Transient electric dichroism studies of the structure of the DNA complex with intercalated drugs. Biochemistry. 1979 Jan 23;18(2):280–288. doi: 10.1021/bi00569a007. [DOI] [PubMed] [Google Scholar]
  11. Jain S. C., Tsai C. C., Sobell H. M. Visualization of drug-nucleic acid interactions at atomic resolution. II. Structure of an ethidium/dinucleoside monophosphate crystalline complex, ethidium:5-iodocytidylyl (3'-5') guanosine. J Mol Biol. 1977 Aug 15;114(3):317–331. doi: 10.1016/0022-2836(77)90253-4. [DOI] [PubMed] [Google Scholar]
  12. Jones C. E., Olson O. M. Sequence-specific DNA-protein interaction: the lac repressor. J Theor Biol. 1977 Jan 21;64(2):323–332. doi: 10.1016/0022-5193(77)90360-5. [DOI] [PubMed] [Google Scholar]
  13. Jovin T. M., Geisler N., Weber K. Amino-terminal fragments of Escherichia coli lac repressor bind to DNA. Nature. 1977 Oct 20;269(5630):668–672. doi: 10.1038/269668a0. [DOI] [PubMed] [Google Scholar]
  14. Kania J., Müller-Hill B. Construction, isolation and implications of repressor-galactosidase - beta-galactosidase hybrid molecules. Eur J Biochem. 1977 Oct 3;79(2):381–386. doi: 10.1111/j.1432-1033.1977.tb11819.x. [DOI] [PubMed] [Google Scholar]
  15. McKay D. B., Pickover C. A., Steitz T. A. Escherichia coli lac repressor is elongated with its operator DNA binding domains located at both ends. J Mol Biol. 1982 Mar 25;156(1):175–183. doi: 10.1016/0022-2836(82)90465-x. [DOI] [PubMed] [Google Scholar]
  16. Nick H., Arndt K., Boschelli F., Jarema M. A., Lillis M., Sadler J., Caruthers M., Lu P. lac repressor-lac operator interaction: NMR observations. Proc Natl Acad Sci U S A. 1982 Jan;79(2):218–222. doi: 10.1073/pnas.79.2.218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ogata R. T., Gilbert W. DNA-binding site of lac repressor probed by dimethylsulfate methylation of lac operator. J Mol Biol. 1979 Aug 25;132(4):709–728. doi: 10.1016/0022-2836(79)90384-x. [DOI] [PubMed] [Google Scholar]
  18. Provencher S. W. A Fourier method for the analysis of exponential decay curves. Biophys J. 1976 Jan;16(1):27–41. doi: 10.1016/S0006-3495(76)85660-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pörschke D. Structure and dynamics of a tryptophanepeptide-polynucleotide complex. Nucleic Acids Res. 1980 Apr 11;8(7):1591–1612. doi: 10.1093/nar/8.7.1591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Stellwagen N. C. Electric birefringence of restriction enzyme fragments of DNA: optical factor and electric polarizability as a function of molecular weight. Biopolymers. 1981 Mar;20(3):399–434. doi: 10.1002/bip.1981.360200302. [DOI] [PubMed] [Google Scholar]
  21. Wells R. D., Hardies S. C., Horn G. T., Klein B., Larson J. E., Neuendorf S. K., Panayotatos N., Patient R. K., Selsing E. RPC-5 column chromatography for the isolation of DNA fragments. Methods Enzymol. 1980;65(1):327–347. doi: 10.1016/s0076-6879(80)65043-5. [DOI] [PubMed] [Google Scholar]

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