Serpins in prokaryotes

Members of the serpin (serine proteinase inhibitor) superfamily have been identified in higher multicellular eukaryotes (plants and animals) and viruses but not in bacteria, archaea, or fungi. Thus, the ancestral serpin and the origin of the serpin inhibitory mechanism remain obscure. In this study we characterize 12 serpin-like sequences in the genomes of prokaryotic organisms, extending this protein family to all major branches of life. Notably, these organisms live in dramatically different environments and some are evolutionarily distantly related. A sequence-based analysis suggests that all 12 serpins are inhibitory. Despite considerable sequence divergence between the proteins, in four of the 12 sequences the region of the serpin that determines proteinase specificity is highly conserved, indicating that these inhibitors are likely to share a common target. Inhibitory serpins are typically prone to polymerization upon heating; thus, the existence of serpins in the moderate thermophilic bacterium Thermobifida fusca, the thermophilic bacterium Thermoanaerobacter tengcongensis, and the hyperthermophilic archaeon Pyrobaculum aerophilum is of particular interest. Using molecular modeling, we predict the means by which heat stability in the latter protein may be achieved without compromising inhibitory activity.     

Glycoproteins in prokaryotes

Rather recently it has become clear that prokaryotes (Archaea and Bacteria) are able to glycosylate proteins. A literature survey revealed the different types of glycoproteins. They include mainly surface layer (S-layer) proteins, flagellins, and polysaccharide-degrading enzymes. Only in a few cases is structural information available. Many different structures have been observed that display much more variation than that observed in eukaryotes. A few studies have given evidence for the function of the prokaryotic glycoprotein glycans. Also from the biosynthetic point of view, information is rather scarce. Due to their different cell structure, prokaryotes have to use mechanisms different from those found in eukaryotes to glycosylate proteins. However, from the fragmented data available for the prokaryotic glycoproteins, similarities with the eukaryotic system can be noticed. Received: 24 February 1997 / Accepted: 13 May 1997     

Genome organization in prokaryotes

Most of the well-characterized prokaryotic genomes consist of double-stranded DNA organized as a single circular chromosome 0.6–10 Mb in length and one or more circular plasmid species of 2 kb-1.7 Mb. The past few years, however, have revealed some major variations in genome organization. In addition, a recent accumulation of data has shown that the location and orientation of the genes and repeated sequences (including prophages and transposons) on and among these elements is not always random. Some of the non-randomness is probably the result of unique historical events; in other cases it reflects selection for the optimization of function.     

Circadian clocks in prokaryotes

Prokaryotes have long been thought incapable of expressing circadian (daily) rhythms. Recently, however, such biological 'clocks' have been discovered in several species of cyanobacteria. These endogenous timekeepers control gene expression on a global level in cyanobacteria. Even in cyanobacterial cultures that are growing with average doubling times more rapid than one per 24 h, the circadian clock controls gene expression and cell division. We have isolated mutants of the cyanobacterial circadian pacemaker and are currently characterizing the loci responsible for their altered period phenotypes.     

Thiamin biosynthesis in prokaryotes

Twelve genes involved in thiamin biosynthesis in prokaryotes have been identified and overexpressed. Of these, six are required for the thiazole biosynthesis (thiFSGH, thiI, and dxs), one is involved in the pyrimidine biosynthesis (thiC), one is required for the linking of the thiazole and the pyrimidine (thiE), and four are kinase genes (thiD, thiM, thiL, and pdxK). The specific reactions catalyzed by ThiEF, Dxs, ThiDM, ThiL, and PdxK have been reconstituted in vitro and ThiS thiocarboxylate has been identified as the sulfur source. The X-ray structures of thiamin phosphate synthase and 5-hydroxyethyl-4-methylthiazole kinase have been completed. The genes coding for the thiamin transport system (thiBPQ) have also been identified. Remaining problems include the cloning and characterization of thiK (thiamin kinase) and the gene(s) involved in the regulation of thiamin biosynthesis. The specific reactions catalyzed by ThiC (pyrimidine formation), and ThiGH and ThiI (thiazole formation) have not yet been identified. Received: 23 August 1998 / Accepted: 16 January 1999     

Insulin-related material in prokaryotes

Abstract We have reported previously on the presence of vertebrate-type peptide hormones in Tetrahymena pyriformis and Escherichia coli . We now have examined other prokaryotes for immunologically detectable insulin-like material. The bacteria studied were E. coli, Acinetobacter calcoaceticus RAG-1, Bordetella pertussis and Halobacteria solinarium; they were grown in defined minimal salt media under varying growth conditions. All these bacteria contain insulin-related material (1–12 pg per g cell wet wt). No insulin immunoactivity was detected in the media prior to inoculation. The content of insulin-related material was not affected by the carbon source used for cell growth. These data are in good agreement with data published previously, and suggest that prokaryotes and unicellular eukaryotes are capable of producing hormone-like material; the function of these peptides, if any, is as yet unknown.     

Silent genes in prokaryotes

Abstract DNA sequence analysis provides excellent evidence for the origin of new genes, encoding new enzyme specificities or isozymes, via gene-duplication. New genes which arise in this way are likely to have arisen via silent gene intermediates. Such 'silent' genes are conceptually distinct from 'cryptic' genes which may also be silent; whereas cryptic genes are thought to be retained due to periodic selection, silent genes would be expected to have only a transient existence in the genome. Only very few of the known inactive genes are possibly (and with varying degrees of likelihood) of the 'silent' type.     

Glutathione analogs in prokaryotes

Background

Oxygen is both essential and toxic to all forms of aerobic life and the chemical versatility and reactivity of thiols play a key role in both aspects. Cysteine thiol groups have key catalytic functions in enzymes but are readily damaged by reactive oxygen species (ROS). Low-molecular-weight thiols provide protective buffers against the hazards of ROS toxicity. Glutathione is the small protective thiol in nearly all eukaryotes but in prokaryotes the situation is far more complex.

Scope of review

This review provides an introduction to the diversity of low-molecular-weight thiol protective systems in bacteria. The topics covered include the limitations of cysteine as a protector, the multiple origins and distribution of glutathione biosynthesis, mycothiol biosynthesis and function in Actinobacteria, recent discoveries involving bacillithiol found in Firmicutes, new insights on the biosynthesis and distribution of ergothioneine, and the potential protective roles played by coenzyme A and other thiols.

Major conclusions

Bacteria have evolved a diverse collection of low-molecular-weight protective thiols to deal with oxygen toxicity and environmental challenges. Our understanding of how many of these thiols are produced and utilized is still at an early stage.

General significance

Extensive diversity existed among prokaryotes prior to evolution of the cyanobacteria and the development of an oxidizing atmosphere. Bacteria that managed to adapt to life under oxygen evolved, or acquired, the ability to produce a variety of small thiols for protection against the hazards of aerobic metabolism. Many pathogenic prokaryotes depend upon novel thiol protection systems that may provide targets for new antibacterial agents. This article is part of a Special Issue entitled Cellular functions of glutathione.     

Photoregulation systems for light-oriented chloroplast movement

Movements of the chloroplasts induced by a directional stimulus of light are found to occur in various plant materials ranging from algae to terrestrial angiosperms. Depending on the fluence rate of light, chloroplasts move toward the area of the maximum light absorption and escape from it, which are named as low- and high-fluence-rate responses, respectively. In most materials the effective wavelengths are exclusively found in the blue-UV region of spectrum, (a) flavin pigment(s) being considered as the photoreceptor, while in a few species of plants phytochrome is involved. The arrangement of chloroplasts as a result of the movement depends on the orientation of the electrical vector of light, which reflects the dichroic orientation of the photoreceptor for perception of light direction. Photosystems involved in these responses, however, are not only for perception of light direction, but also for realization of the movement and for holding of the chloroplasts in the reached site. Possible interactions and dual roles of photosystems in regulation of chloroplast movement are discussed.