Catabolite repression of the lac operon: effect of operator-constitutive mutations

Summary We have constructed and tested three lac diploid strains in an attempt to show whether operator-constitutive mutations relieve catabolite repression of the lac operon. Each of these carries a different operator mutation on the chromosome, and all three have the genotype I⁺P⁺O^cZ⁺Y^-polar/Flac I⁺P⁺O⁺Z^delY⁺A⁺. When these strains were grown in medium containing glucose plus gluconate, synthesis of -galactosidase (directed by a gene cis to a mutant operator) and of thiogalactoside transacetylase (directed by a gene cis to an intact operator) suffered equal catabolite repression. We conclude that the operator-constitutive mutations have no effect on catabolite repression. Since it has been shown in analogous experiments that all promoter mutations tested do alleviate catabolite repression, these results are consistent with the view that the operator and promoter are functionally distinct.     

Transient repression of the lac operon - the effect of a lac promoter deletion

Experiments have been done to show whether the lac promoter delection L1, which partly alleviates catabolite repression, also affects transient repression of lac. In stain L1/F'M15 all of the beta-galactosidase is synthesized from a chromosomal gene cis to L1, whereas 98% of the thiogalactosidase transacetylase is synthesized from an episomal gene cis to an intact i-p-o region. The addition of glucose to induced cultures of strain L1/F'M15 growing in glycerol medium caused extensive transient repression of transacetylase but almost no transient repression of beta-galactosidase. In control experiments with a diploid stain of genotype p(+)z(+)a(-)/F'p(+)z(-)a(+) the two enzymes suffered equal transient repression. Thus L1 substantially relieves transient repression.     

The lac operon galactoside acetyltransferase

Of the proteins encoded by the three structural genes of the lac operon, the galactoside acetyltransferase (thiogalactoside transacetylase, LacA, GAT) encoded by lacA is the only protein whose biological role remains in doubt. Here, we briefly note the classical literature that led to the identification and initial characterization of GAT, and focus on more recent results which have revealed its chemical mechanism of action and its membership in a large superfamily of structurally similar acyltransferases. The structural and sequence similarities of several members of this superfamily confirm the original claim for GAT as a CoA-dependent acetyltransferase specific for the 6-hydroxyl group of certain pyranosides, but do not yet point to the identity of the natural substrate(s) of the enzyme.     

Paradoxical effect of weak inducers on the lac operon of Escherichia coli

Previously, we reported the existence of a group of compounds whose function in the regulation of the lac operon was "paradoxical" in that they acted as either inducers or repressors depending on the circumstances. We now show that this group of compounds does not repress the lac operon by catabolite repression, transient repression, or by preventing the uptake of inducers. A model is presented which shows that "paradoxical" behavior is to be expected if a weak inducer is present at a concentration that is high relative to its binding affinity for the regulatory macromolecule. This model depends on the assumptions that the regulatory macromolecule is an allosteric protein which undergoes a transition between two conformational states and that the rate of enzyme synthesis depends on the fraction of protein molecules in each state. The previous observations on the responses of lac regulatory mutants to weak inducers have been extended to a series of such mutants. Weak inducers repress beta-galactosidase synthesis in several i(-) mutants. When this happens, enzyme synthesis can be reinduced by using a strong inducer such as isopropyl-beta-d-thiogalactoside. These compounds induce operator constitutives and the i(t) mutant more easily than they induce a wild-type strain.     

Understanding the placebo effect from an evolutionary perspective

A placebo is a treatment which is not effective through its direct action on the body, but works because of its effect on the patient's beliefs. From an evolutionary perspective, it is initially puzzling why, if people are capable of recovering, they need a placebo to do so. Based on an argument put forward by Humphrey [Great expectations: the evolutionary psychology of faith-healing and the placebo effect. In: Humphrey, N (2002). The mind made flesh. Oxford University Press, Oxford. 255–285], we present simple mathematical models of the placebo effect that involve a trade-off between the costs and benefits of allocating resources to a current problem. These models show why the effect occurs and how its magnitude and timing can depend on different factors. We identify a particular aspect of belief which may govern the effect and conclude that a deeper understanding of why the placebo effect exists may allow it to be invoked more easily in the future.     

The mast cell: an evolutionary perspective

This review article is an attempt to trace the evolution of mast cells (MCs). These immune cells have been identified in all vertebrate classes as single‐lobed cells containing variable amounts of membrane‐bound secretory granules which store a large series of mediators, namely histamine, proteases, cytokines and growth factors. Other MC features, at least in mammals, are the c‐kit receptor for the stem cell factor and the high‐affinity receptor, FcεRI, for immunoglobulin E (IgE). The c‐kit receptor also has been identified in fish MCs. The FcεRI receptor seems to be a more recent acquisition in MC phylogenesis given that IgE originated in mammalian species. Tryptase and histamine have also been recognized in MCs of teleost fish. Thus, a cell population with the overall characteristics of higher vertebrate MCs is identifiable in the most evolutionarily advanced fish species. Two potential MC progenitors have been identified in ascidians (urochordates which appeared approximately 500 million years ago): the basophil/MC‐like granular haemocyte and the test cell. Both contain histamine and heparin, and provide defensive functions. Some granular haemocytes in Arthropoda also closely approximate the ultrastructure of modern MCs. The origin of MCs is probably to be found in a leukocyte ancestor operating in the context of a primitive local innate immunity and involved in phagocytic and killing activity against pathogens. From this type of defensive cell, the MC phylogenetic progenitor evolved into a tissue regulatory and remodelling cell, which was incorporated into the networks of recombinase activating genes (RAG)‐mediated adaptive immunity in the Cambrian era, some 550 million years ago. Early MCs probably appeared in the last common ancestor we shared with hagfish, lamprey and sharks about 450‐500 million years ago.     

The hygiene hypothesis: an evolutionary perspective

The hygiene hypothesis relies on the assumption that humans have adapted to a pathogen-rich environment that no longer exists in industrialized societies. Recent advances in molecular immunology and population genetics allow deeper insight into the evolution and co-evolution of host–pathogen interactions and, therefore, into the foundations of the hygiene hypothesis.     

Marriage: an evolutionary perspective

Marriage is universal, and pair bonding is found in other species too with highly dependent young. So marriage functions as a reproductive social arrangement that traditionally involved the extended family. The sexes are not identical in their biological contributions to children's survival, so they seek somewhat different attributes in a mate. Men seek a young, attractive, sexually faithful bride. Women seek a man who is older, taller, and (as in many other species) socially dominant. Both sexes prefer a kind, healthy, attractive, similar mate who is emotionally attached to them. A spouse who fails to maintain sufficiently high mate value is vulnerable to divorce. Infertility and sexual dissatisfaction predict divorce, as does death of a child, but the more children, the stabler the marriage. Cross-cultural data suggest that cruel or subdominant men (e.g., poor providers) and unfaithful women are prone to divorce. Marriages in which the wife dominates the husband in economic contributions, nonverbal behavior, and decision making tend to be less satisfying. In societies in which wives are economically independent of husbands, divorce rates are high. As women's economic power has risen with industrialization, divorce rates have climbed. Economic and fitness considerations also help explain cultural differences in polygyny, age at marriage, arranged marriage, concern with the bride's sexual chastity, and marriage ceremonies. Other factors also affect marital dynamics, such as state subsidies to families, the sex ratio, and influence of the couple's parents.     

Osteoporosis: an evolutionary perspective

Increased life expectancy has led to an overall aging of the population and greater numbers of elderly people. Therefore, the number of people with osteoporosis has increased substantially, accompanied with an epidemic of hip fractures. Osteoporosis is an age-related systemic condition that naturally occurs, among mammals, only in humans. Osteoporosis is known to be highly heritable. However, assuming a genetic determinant for this post-reproductive disease to be transmitted from one generation to the next is counter-intuitive, based on the principles of human evolution, I will attempt to provide an explanation of the phenomenon from the point of view of evolution, selection, and changed environment in humans, which contributed to human longevity, while on other hand, contribute to diseases of civilization, including osteoporosis. There is a need to delve into evolution of human species in search for adaptive patterns to a specific environment that humans are operating in the last couple of millennia, to clarify whether “good” and “bad” genes exist, and how to find and correct them. The answer to the above questions will help us to identify causes of the current epidemic of osteoporosis and to pin-point a tailored treatment.     

Structure of the lac operon galactoside acetyltransferase

The galactoside acetyltransferase (thiogalactoside transacetylase) of Escherichia coli (GAT, LacA, EC 2.3.1.18) is a gene product of the classical lac operon. GAT may assist cellular detoxification by acetylating nonmetabolizable pyranosides, thereby preventing their reentry into the cell. The structure of GAT has been solved in binary complexes with acetyl-CoA or CoA and in ternary complexes with CoA and the nonphysiological acceptor substrates isopropyl beta-D-thiogalactoside (IPTG) or p-nitrophenyl beta-D-galactopyranoside (PNPbetaGal). A hydrophobic cleft that binds the thioisopropyl and p-nitrophenyl aglycones of IPTG and PNPbetaGal may discriminate against substrates with hydrophilic substituents at this position, such as lactose, or inducers of the lac operon. An extended loop projecting from the left-handed parallel beta helix domain contributes His115, which is in position to facilitate attack of the C6-hydroxyl group of the substrate on the thioester.     

Hyphal morphogenesis: an evolutionary perspective

Two modes of cellular morphogenesis predominate within the fungal kingdom; yeast growth and hyphal growth. The availability of complete genome sequences that span the kingdom has made possible the use of comparative approaches that address important questions regarding the evolution of these growth modes. These comparisons have also emphasized the point that not all hyphae are the same despite outward appearances. Topics considered here include the origins of hyphal growth, as well as the potential causes of and the consequences resulting from the loss of hyphal growth in yeast lineages. The mechanisms that enable distinct morphological outputs (i.e., yeast vs. hyphae) using an essentially identical inventory of gene products are also considered. Finally, processes implicated in the regulation of hyphal tip complexes are addressed from an evolutionary perspective.     

Prions: an evolutionary perspective.

Studies in both prion-due diseases in mammals and some non-Mendelian hereditary processes in yeasts have demonstrated that certain proteins are able to transmit structural information and self-replication. This induces the corresponding conformational changes in other proteins with identical or similar sequences. This ability of proteins may have been very useful during prebiotic chemical evolution, prior to the establishment of the genetic code. During this stage, proteins (proteinoids) must have molded and selected their structural folding units through direct interaction with the environment. The proteinoids that acquired the ability to propagate their conformations (which we refer to as conformons) would have acted as reservoirs and transmitters of a given structural information and hence could have acted as selectors for conformational changes. Despite the great advantage that arose from the establishment of the genetic code, the ability to propagate conformational changes did not necessarily disappear. Depending on the degree of involvement of this capacity in biological evolution, we propose two not mutually exclusive hypotheses: (i) extant prions could be an atavism of ancestral conformons, which would have co-evolved with cells, and (ii) the evolution of conformons would have produced cellular proteins, able to transmit structural information, and, in some cases, participating in certain processes of regulation and epigenesis. Therefore, prions could also be seen as conformons of a conventional infectious agent (or one that co-evolved with it independently) that, after a longer or shorter adaptive period, would have interacted with conformons from the host cells.     

The Chikungunya threat: an ecological and evolutionary perspective

Chikungunya virus (CHIKV) is an emerging mosquito-borne alphavirus. Although primarily African and zoonotic, it is known chiefly for its non-African large urban outbreaks during which it is transmitted by the same vectors as those of Dengue viruses. Unlike Dengue viruses, CHIKV displays a re-emergence pattern that closely depends on long-distance migrations including recent re-immigrations from African (putatively zoonotic) sources. Genus-based differences also emerged when comparing the evolution of Dengue-related (Flaviviruses) and of CHIKV-related (Alphaviruses) arboviruses. In this review, we discuss current information on CHIKV genetics, ecology and human infection. Further investigations on African CHIKV ecology and the differences between Flavivirus and Alphavirus members in adaptive changes and evolutionary constraints are likely to help delineate the potential of further CHIKV (re-)emergence.     

Catabolite repression of the lac operon. Effect of mutations in the lac promoter

1. Several lac diploid strains of Escherichia coli were constructed and tested to discover whether mutations in the lac promoter alleviate catabolite repression. 2. In each of these diploids the chromosome carries one of the promoter mutations, L8, L29 or L1; so that the rate of synthesis of the enzymes of the lac operon is only 2-6% of the fully induced wild-type. Each diploid harbours the episome F'lacM15 that specifies the synthesis of thiogalactoside transacetylase under the control of intact regulator, promoter and operator regions, but has a deletion in the structural gene for beta-galactosidase. In each diploid more than 90% of the thiogalactoside transacetylase is synthesized from the episome, and 100% of the beta-galactosidase is synthesized from the chromosome, and comparison of the extent of catabolite repression that the two enzymes suffered indicated whether the chromosomal promoter mutation relieves catabolite repression. 3. In the strains in which the promoter carries either of the point mutations L8 or L29 the enzymes were equally repressed, suggesting that neither L8 nor L29 affects catabolite repression. 4. In a diploid strain harbouring the same episome but carrying deletion L1 on the chromosome, synthesis of beta-galactosidase suffered much less repression than that of thiogalactoside transacetylase. 5. In a diploid strain in which the chromosome carries L1 and also a second mutation that increases the rate of expression of lac to that permitted by L8 or L29, the synthesis of beta-galactosidase again suffered much less repression than the synthesis of thiogalactoside transacetylase. 6. The effect of L1 (which deletes the boundary between the i gene and the lac promoter) is ascribed to its bringing the expression of lac under the control of the promoter of the i gene. 7. Even in strains carrying L1, some catabolite repression persists; this is not due to a trans effect from the episome since it occurs equally in a haploid strain with L1.