Escherichia coli Lactose Operon

Abstract

The study of Escherichia coli lactose operon laid the foundation of modern molecular biology. It contributed to the elaboration of the concept of genetic regulation, proposed by Jacob and Monod almost 50 years ago, a model which survives essentially unchanged. The operon structure, consisting of structural and regulatory genes has been elaborated and their regulatory response to small molecules, such as inducer, glucose and cyclic AMP, have been elucidated. Gene regulation of the lactose operon led to the discovery of messenger ribonucleic acid (mRNA), to the identification of the Lac repressor and to the development of the theory of allostery. The lactose genes and its derivatives provide tools to a wide range of current applications in many fields of biology; they are the most commonly used reporter genes in the analysis of developmentally regulated systems, in the study of mutation frequencies and in functional genomics.

Keywords: complementation; fusion proteins; repressor; gene regulation; cyclic AMP; allostery

Figure 1.

The lac structural genes.

Figure 2.

Three‐dimensional structure of β‐galactosidase. (a) Ribbon representation of the β‐galactosidase tetramer showing the largest face of the molecule. Contacts between red/green and blue/yellow dimers form the long interface. Contacts between the red/yellow and blue/green dimers form the activating interface. Formation of the tetrameric particle results in two deep clefts that run across opposite faces of the molecule. Each contain two active sites. (b) Ribbon diagram of the blue/green dimer viewed down the molecular 2‐fold axis, showing the composition of the activating interface. Residues 1–50 from each chain, which form the α‐complementation region (see text), are shown in red. The interface includes contacts between the respective complementation peptides, between two helices from the respective monomers that pack together to form a four‐helix bundle, and between an extended loop (residues 272–288) from each monomer that reaches across the interface and extends into the active site region of the neighbouring monomer stabilizing the active site structure. (c) Stereo ribbon diagram of the β‐galactosidase monomer showing the domain organization of the chain. Residues corresponding to successive domains are coloured in successive spectral colours. Reprinted from Jacobson RH, Zhang XJ, DuBose RF and Matthews BW (1994) Three‐dimensional structure of β‐galactosidase of E. coli. Nature369;761–766, with permission.

Figure 3.

The ‘PaJaMo’ experiment. A male (Hfr) lac i+z+ strain was conjugated with a female (F) lac iz strain in the absence of inducer. At the time indicated, inducer was added to one of the cultures, whereas the other one received no addition. β‐Galactosidase activity was measured as a function of time. Adapted from Pardee AB, Jacob F and Monod J (1959) The genetic control and cytoplasmic expression of ‘inducibility’ in the synthesis of β‐galactosidase by E. coli. Journal of Molecular Biology1; 165–178, with permission.

Figure 4.

The crystal structure of the Lac repressor–DNA complex constructed from the available Protein Data Bank structures (Lewis et al., 1996) by modelling procedures (Balaeff et al., 2004). In the V‐shaped tetrameric Lac repressor each of the two dimers (drawn as purple protein cartoon) binds with high specificity to a 21‐base pair operator DNA fragment (drawn as blue tubes and red spheres) through the N‐terminal 62 residue‐long headpiece (drawn in green). Dimer–dimer assembly occurs via a compact four‐helical bundle formed by 18 C‐terminal residues from each subunit (drawn as orange tubes). Adapted from Balaeff A, Mahadevan L and Schulten K (2004) Structural basis for cooperative DNA binding by CAP and Lac repressor. Structure12; 123–132, with permission from Elsevier.

Figure 5.

Diagram of the lactose operon in the repressed (a) and induced (b) states. Synthesis of the lactose operon proteins, genetically determined by the structural genes (lacZ, lacY and lacA), is blocked by the LacI repressor synthesized by the regulator gene, lacI. The operator (O) is the site of specific interaction with the repressor. The repressor can be inactivated by the inducer, thus allowing transcription to take place at the promoter. Inherent to the operon model is the assumption that transfer of genetic information from gene to protein involves a short‐lived mRNA. Reprinted with permission from Jacob F and Monod J (1961) Genetic regulatory mechanisms in the synthesis of proteins. Journal of Molecular Biology3: 318–356. Copyright © 1961 Academic Press. Drawing courtesy of Jean‐Marc Ghigo.

Figure 6.

Schematic structure of the CAP–DNA complex. For the DNA, sharply bent by CAP, the bases and backbone are shown in red. The protein is represented as a ribbon diagram (blue) with two cAMP molecules (red) placed to indicate the binding sites on each subunit of CAP. Reprinted from Parkinson G, Gunasekera A Vojtechovsky J et al. (1996) Aromatic hydrogen bond in sequence‐specific protein DNA recognition. Nature Structural Biology3: 837–841, with permission from Nature Publishing Group. Figure courtesy of Richard H. Ebright.

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Further Reading

Balaeff A, Mahadevan L and Schulten K (2004) Structural basis for cooperative DNA binding by CAP and Lac repressor. Structure 12: 123–132.

Beckwith JR and Zipser D (eds) (1970) The Lactose Operon. Cold Spring Harbor: Cold Spring Harbor Laboratory Press.

Fowler A and Zabin I (1978) Amino acid sequence of β‐galactosidase. Journal of Biological Chemistry 253: 5521–5525.

Jacob F and Monod J (1961) Genetic regulatory mechanisms in the synthesis of proteins. Journal of Molecular Biology 3: 318–356.

Jacob F, Ullmann A and Monod J (1965) Délétions fusionnant l'opéron lactose et un opéron purine chez E. coli. Journal of Molecular Biology 31: 704–719.

Jacobson RH, Zhang XJ, DuBose RF and Matthews BW (1994) Three‐dimensional structure of β‐galactosidase of E. coli. Nature 369: 761–766.

Kolb A, Busby S, Buc H, Garges S and Adhya S (1993) Transcriptional activation by cAMP and its receptor protein. Annual Review of Biochemistry 62: 749–795.

Lewis M, Chang G, Horton NC et al. (1996) Structure of the E. coli lactose operon repressor and its complexes with DNA and inducer. Science 271: 1247–1254.

Lwoff A and Ullmann A (eds) (1978) Selected Papers in Molecular Biology by Jacques Monod. New York: Academic Press.

Lwoff A and Ullmann A (eds) (1979) Origins of Molecular Biology. A Tribute to Jacques Monod. New York: Academic Press.

Monod J (1945) Sur la nature du phénomène de diauxie. Annales de l'Institut Pasteur Paris 71: 37–40.

Monod J (1966) From enzymatic adaptation to allosteric transitions. Science 154: 475–483.

Müller‐Hill B (1996) The lac Operon. A Short History of a Genetic Paradigm. Berlin: Walter de Gruyter.

Pace HC, Kercher MA, Lu P et al. (1997) Lac repressor genetic map in real space. Trends in Biochemical Sciences 22: 334–339.

Pardee AB, Jacob F and Monod J (1959) The genetic control and cytoplasmic expression of ‘inducibility’ in the synthesis of β‐galactosidase by E. coli. Journal of Molecular Biology 1: 165–178.

Rossi FMV, Blakely BT and Blau HM (2000) Interaction blues: protein interactions monitored in live mammalian cells by β‐galactosidase complementation. Trends in Cell Biology 10: 119–122.

Ullmann A (1992) Complementation in β‐galactosidase: from protein structure to genetic engineering. BioEssays 14: 201–205.

Ullmann A (ed.) (2003) Origins of Molecular Biology. A Tribute to Jacques Monod. Washington, DC: ASM press.

Ullmann A and Danchin A (1983) Role of cyclic AMP in bacteria. Advances in Cyclic Nucleotide Research 15: 1–53.

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How to Cite close
Ullmann, Agnes(Mar 2009) Escherichia coli Lactose Operon. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0000849.pub2]