A Legionella pneumophila Peptidyl-Prolyl cis-trans Isomerase Present in Culture Supernatants Is Necessary for Optimal Growth at Low Temperatures

Several Legionella pneumophila proteins were highly expressed in low-temperature supernatants. One of these proteins was the peptidyl-prolyl isomerase PpiB. Mutants lacking ppiB exhibited reduced growth at 17°C. Since PpiB lacked a signal sequence and was present in 17°C supernatants of type II and type IV secretion mutants, this protein may be secreted by a novel mechanism.     

Root Growth and Carbohydrate Metabolism at Low Temperatures

A study of carbohydrate metabolism in the roots of pea and maizeshows that the differing ability of these two species to growat low temperatures is associated with the maintenance of adequatesugar supplies to the root tip. In the maize root reducing theambient temperature to 2 °C causes a sharp and continuingfall in the soluble sugar content of the growing tip. A similartreatment with pea roots causes only a temporary reduction insugar content lasting no more than 24 h. The fall in root sugarsin maize is accompanied by a reduction in respiration rate andthe cessation of growth. During the periods of sugar shortagecaused by low temperatures both respiration and growth can bestimulated in root tips by a supply of exogenous glucose. Pearoots also show an additional ability to adapt to low temperaturesby lowering the K^m value for invertase after pretreatment ata low temperature. This effect is not seen in maize.     

Growth, respiration and survival of Legionella pneumophila at high temperatures

The optimum temperature for multiplication of legionella strains in culture media is around 37°C. The effect of high temperatures on the growth of strains isolated from various environments is poorly known. We studied the growth (cell multiplication, respiration) of clinical and environmental Legionella pneumophila strains in liquid media at intervals of 0.5°C in the temperature range from 41.6 to 51.6°C using a temperature gradient incubator. Cell multiplication and CO₂ production decreased markedly with all the strains at temperatures above 44–45°C. CO₂ continued to be produced up to 51.6C even if cell multiplication generally stopped at around 48.4–50.0C. Thus, legionella retained its metabolic activity beyond the maximum temperature for cell multiplication. The CO₂ production per bacterial cell (metabolic quotient, qCO₂) increased with increasing temperature up to 45°C, whereafter it decreased, the turning point being almost at the same at which the rate of cell multiplication decreased. The difference in qCO₂ between the strains may reflect their different physiological capacities for tolerating high temperatures.     

Importance of Type II Secretion for Survival of Legionella pneumophila in Tap Water and in Amoebae at Low Temperatures

Legionella pneumophila type II secretion mutants showed reduced survival in both tap water at 4 to 17°C and aquatic amoebae at 22 to 25°C. Wild-type supernatants stimulated the growth of these mutants, indicating that secreted factors promote low-temperature survival. There was a correlation between low-temperature survival and secretion function when 12 additional Legionella species were examined.     

A Carbon 14 Study of Metabolism in Tall Fescue Seedlings Acclimating to Growth at Low Temperatures

The youngest fully expanded leaves of young plants of tall fescue(Festuca arundinacea Schreb. cv. S 170) were allowed to assimilate¹⁴CO₂ either (a) at the fifth leaf stage immediately beforetransfer from 17/14?C to 7/4?C (non-acclimated), or (b) aftertransfer from 17/14?C to 7/4?C at the fifth leaf stage, andfurther development in 7/4?C of one or two more leaves (acclimationfor one or two plastochrons). Controls were maintained in 17/14?C and allowed to assimilate ¹⁴CO₂ at the corresponding (fifth,sixth, or seventh) leaf stages. Allocation of 14C amongst sinksand biochemical fractions was analysed during a subsequent periodof growth at 7/4 ?C (cold) or 17/14 ?C (control). Allocationof assimilate to growing parts of the shoot was less than controlsin the non-acclimated plants transferred to the cold and morethan controls in plants acclimated for one plastochron. Afterthe imbalance induced initially by transfer from 17/14?C to7/4 ?C, acclimation for one or two plastochrons brought theallocation amongst sinks and amongst biochemical fates closerto the balance existing in controls. The main shoot and sidetillers differed in the time during acclimation when they becamemore like the controls. The percentage of assimilate in thesoluble carbohydrate fraction of non-acclimated plants and ofplants acclimated for one plastochron, was higher than in controls.The percentage of assimilate in the cell wall fraction was lowerin non-acclimated plants than in controls but the differencefrom controls after two plastochrons acclimation again was less.During the first plastochron after transfer to cold, diversionof assimilate to a reserve pool in sinks could be a factor limitingcell wall synthesis and growth and this limitation may be relievedwhen the reserve pool is ‘full’. Low temperaturereduced the percentage of assimilate in the protein fractionof growing parts of the shoot and the difference was found evenafter two plastochrons acclimation. Key words: Low temperature, metabolism, tall fescue (Festuca arundinacea Schreb. cv. S. 170)     

Glucose Metabolism in Legionella pneumophila: Dependence on the Entner-Doudoroff Pathway and Connection with Intracellular Bacterial Growth

Glucose metabolism in Legionella pneumophila was studied by focusing on the Entner-Doudoroff (ED) pathway with a combined genetic and biochemical approach. The bacterium utilized exogenous glucose for synthesis of acid-insoluble cell components but manifested no discernible increase in the growth rate. Assays with permeabilized cell preparations revealed the activities of three enzymes involved in the pathway, i.e., glucokinase, phosphogluconate dehydratase, and 2-dehydro-3-deoxy-phosphogluconate aldolase, presumed to be encoded by the glk, edd, and eda genes, respectively. Gene-disrupted mutants for the three genes and the ywtG gene encoding a putative sugar transporter were devoid of the ability to metabolize exogenous glucose, indicating that the pathway is almost exclusively responsible for glucose metabolism and that the ywtG gene product is the glucose transporter. It was also established that these four genes formed part of an operon in which the gene order was edd-glk-eda-ywtG, as predicted by genomic information. Intriguingly, while the mutants exhibited no appreciable change in growth characteristics in vitro, they were defective in multiplication within eukaryotic cells, strongly indicating that the ED pathway must be functional for the intracellular growth of the bacterium to occur. Curiously, while the deficient glucose metabolism of the ywtG mutant was successfully complemented by the ywtG⁺ gene supplied in trans via plasmid, its defect in intracellular growth was not. However, the latter defect was also manifested in wild-type cells when a plasmid carrying the mutant ywtG gene was introduced. This phenomenon, resembling so-called dominant negativity, awaits further investigation.The Gram-negative bacterium Legionella pneumophila is a facultative intracellular parasite and a clinically important pathogen of human beings. In the natural habitat, it replicates within free-living amoebae in the environment (6). However, once inhaled into humans as contaminated aerosol, it survives and replicates in alveolar macrophages and causes severe pneumonia, Legionnaires'' disease (13).It is generally believed that L. pneumophila utilizes amino acids as carbon and energy sources while it neither ferments nor oxidizes carbohydrates (7). Thus, the formulation of chemically defined media (8, 18, 22) soon led to the discovery that a certain combination of amino acids is sufficient to support the growth of L. pneumophila (21). Almost at the same time, it was also noted that glutamate serves as the principal energy source (23) whereas glucose has no effect on the growth of the bacterium (18, 22). However, despite this widely held notion, there are a few reports that present evidence that glucose is actually metabolized by L. pneumophila, mainly through the Entner-Doudoroff (ED) and/or pentose phosphate pathway (21, 23). In accordance with these observations, the analysis of the genome structure of the bacterium identified the complete set of genes related to the ED pathway ({"type":"entrez-nucleotide","attrs":{"text":"NC_002942","term_id":"52840256","term_text":"NC_002942"}}NC_002942), but the physiological significance of this phenomenon remained unclear. Recently, it has been shown that the expression of the L. pneumophila genes involved in the ED pathway is upregulated within amoebic cells (2). Furthermore, Salmonella enterica serovar Typhimurium, another facultative intracellular pathogen, is known to use gluconate and related carbohydrates by the ED pathway when growing within macrophages (5).In the present work, we tried to clarify the physiological roles of the ED pathway (Fig. ​(Fig.1A)1A) more fully by genetic means, i.e., the construction and characterization of L. pneumophila mutants in which genes essential to the ED pathway are inactive due to insertion mutations. Our first aim was to estimate the degree to which the pathway contributes to glucose metabolism in the bacterium. The results described here clearly showed that the dependence of glucose metabolism on the pathway was nearly complete, thus excluding the possibility of significant contributions by the glycolytic and pentose phosphate pathways. Our second aim was to know whether an active ED pathway was required for intracellular growth of L. pneumophila, as suggested by data obtained with amebic cells (2). In this respect, our results seem to indicate that the functional ED pathway is actually a requirement.Open in a separate windowFIG. 1.(A) The Entner-Doudoroff pathway. The genes zwf, pgl, edd, glk, and eda encode glucose-6-phosphate dehydrogenase, 6-phosphogluconolactonase, phosphogluconate dehydratase, glucokinase, and 2-dehydro-3-deoxy-phosphogluconate aldolase, respectively. (B) Schematic diagram of the DNA fragments used. The large arrows represent genes relevant to this study; vertical dashed lines, ends of coding regions; inverted triangles, sites for insertion of the Km^r cassette; thick lines, DNA fragments for preparation of gene-disrupted mutants; thin lines, DNA inserts in plasmids pMMB207eda (a) and pMMB207ywtG (b) used in complementation; open circles, DNA segments providing a promoter and a ribosome-binding site; dotted lines, probes for Northern hybridization; thin dotted arrow, cDNA synthesized by RT-PCR with primer RT-0; dashed lines, PCR products made on the RT-PCR product as a template with primers RT1 through RT4. For primers and other details, see Materials and Methods and Table S1 in the supplemental material.     

Effect of Low Temperatures on Growth, Structure, and Metabolism of Campylobacter coli SP10

The effect of low temperatures on the survival, structure, and metabolism of Campylobacter coli SP10, a virulent strain, was investigated. C. coli became nonculturable rapidly at 20 and 10°C and slightly later at 4°C. Incubation in a microaerobic atmosphere improved survival, but after day 8, campylobacters were detectable by direct-count procedures only. The increase in the number of coccoid cells was most pronounced at 37°C but also was noticeable at 20 and 10°C. Two forms of coccoid cells were seen electron microscopically, but only one (20 and 10°C) seemed to be a degenerative form. The flagella were shorter at 20 and 10°C, a result which correlates well with the observed slight changes in the 62-kDa protein band. The fatty acid composition of bacterial cells was influenced significantly by low temperatures. An increase in the short-chain and unsaturated acids was noted; above all, a drastic increase in C_19:0 cyc at 20°C with a concomitant decrease in C_18:1 trans9,cis11 was seen. The concentrations of excreted metabolites were analyzed to obtain information on metabolic activity. Depending on the magnitude of the temperature downshift, the production of organic acids decreased, but it was always observable after a temperature-specific lag phase and regardless of ability to be cultured. Under optimal conditions, succinate, lactate, and acetate were the main metabolites, other acids being of less importance. The pattern changed significantly at lower temperatures. Succinate was never detected at 20°C and was only occasionally detected at 10 and 4°C. At the same time, fumarate concentrations, which are normally not detectable at 37°C, were highest at 20°C and reduced at 10 and 4°C. Inactivation of fumarate reductase was considered to be a possible explanation.     

Growth of Bacteria Degrading Naphthalene and Salicylate at Low Temperatures

A total of 58 bacterial strains degrading naphthalene and salicylate were isolated from soil samples polluted with oil products, collected in different regions of Russia during winter and summer. The isolates were assessed for their ability to grow at low temperatures (4, 8, and 15°C); bacteria growing at 4°C in the presence of naphthalene or salicylate accounted for 65 and 53%, respectively, of the strains isolated. The strains differed in the temperature dependence of their growth rates. It was demonstrated that the type of expression of the Nah⁺ phenotype at low temperatures depended on the combination of host bacterium and plasmid.     

An FT-IR Study of DNA and RNA Conformational Transitions at Low Temperatures

Abstract

Fourier Transform Infrared (FT-IR) spectra of solid samples of DNA and RNA obtained from freeze-drying at solid CO₂ and liquid nitrogen temperatures, have been recorded and correlation between the conformational transitions and spectral changes is proposed. It is concluded that an equilibrium exists between A, B and Z conformations at low temperatures for the DNA molecule, which is temperature dependent, whereas the RNA molecule exhibits only the A conformation. The results have been compared with the metal-adducts of DNA and RNA, where one of the conformations is predominant.

Marker infrared bands for the B conformer have been found to be the strong band at 825 cm^?1 (sugar conformer mode) and a band with medium intensity at 690 cm^?1 (guanine breathing mode). The A conformation showed characteristic bands at 810 and 675 cm^?1. The B to Z conformational transition was characterized by the strong absorption bands near 820-810 cm^?1 and at 665-600 cm^?1.     

Effect of Low Temperatures on Growth,Structure, and Metabolism of Campylobacter coli SP10

The effect of low temperatures on the survival, structure, and metabolism of Campylobacter coli SP10, a virulent strain, was investigated. C. coli became nonculturable rapidly at 20 and 10°C and slightly later at 4°C. Incubation in a microaerobic atmosphere improved survival, but after day 8, campylobacters were detectable by direct-count procedures only. The increase in the number of coccoid cells was most pronounced at 37°C but also was noticeable at 20 and 10°C. Two forms of coccoid cells were seen electron microscopically, but only one (20 and 10°C) seemed to be a degenerative form. The flagella were shorter at 20 and 10°C, a result which correlates well with the observed slight changes in the 62-kDa protein band. The fatty acid composition of bacterial cells was influenced significantly by low temperatures. An increase in the short-chain and unsaturated acids was noted; above all, a drastic increase in C_19:0 cyc at 20°C with a concomitant decrease in C_18:1 trans9,cis11 was seen. The concentrations of excreted metabolites were analyzed to obtain information on metabolic activity. Depending on the magnitude of the temperature downshift, the production of organic acids decreased, but it was always observable after a temperature-specific lag phase and regardless of ability to be cultured. Under optimal conditions, succinate, lactate, and acetate were the main metabolites, other acids being of less importance. The pattern changed significantly at lower temperatures. Succinate was never detected at 20°C and was only occasionally detected at 10 and 4°C. At the same time, fumarate concentrations, which are normally not detectable at 37°C, were highest at 20°C and reduced at 10 and 4°C. Inactivation of fumarate reductase was considered to be a possible explanation.

Campylobacters are playing an increasingly important role in gastrointestinal disease all over the world, and in some countries they are even more frequent than salmonellae. In industrialized countries, the annual incidence of enteritis caused by campylobacters is estimated to be 1%. In underdeveloped countries it is much higher, affects mainly small children and, in contrast to the situation in developed countries, is caused more often by Campylobacter coli than by C. jejuni (12, 43, 44). The natural reservoir of campylobacters is the intestine of warm-blooded animals; 24 to 82% of examined chicken populations, 58 to 95% of Scandinavian pigs, and 23% of slaughter cattle were campylobacter positive, to name only a few animal species (1, 9, 19, 41). Campylobacters, like other intestinal pathogens, are released into the environment via the feces of infected animals or humans. On average, raw sewage contains log 2 to 4 CFU of campylobacters per 100 ml, but concentrations can rise significantly if abattoirs or chicken farms are connected to the sewer system (18). Treated sewage may contain campylobacters at log 1 to 2 CFU/100 ml, and fresh, undisinfected sewage sludge, which is used as a fertilizer in some countries, may contain campylobacters at up to log 6 CFU/100 ml (2, 17). Sewage discharge into surface water and agricultural runoff can thus lead to the contamination of bathing water areas and drinking water reservoirs and can pose a significant health risk for the population. Not surprisingly, large waterborne outbreaks of campylobacter-induced enteritis have been reported (31, 45, 48).Various surface waters have been examined in the past, and 25 to 95% were shown to be campylobacter positive, with campylobacter concentrations ranging from log 0.04 to log 2 CFU/100 ml. Detection rates were higher during the winter months, suggesting enhanced survival at low temperatures, although, apart from starvation, temperature downshift is one of the greatest stress factors that bacteria experience when they are released into the environment. While psychrophilic and psychrotrophic bacteria have adapted evolutionarily to life at low temperatures, human pathogens, being mesophilic bacteria, can be severely inhibited. In nutrient-rich media, reactions such as intracellular (p)ppGpp (guanosine 5′-triphosphate-3′-diphosphate and guanosine 5′-diphosphate-3′-diphosphate) synthesis or production of cold shock proteins have been shown to be dependent on the temperature shift (22). Changes in the composition of the cell membrane and inhibition of enzyme activities and membrane transport systems are further conceivable reactions of mesophilic bacteria to temperature downshift (29). To study the effect of low temperatures on campylobacters, a virulent C. coli strain was subjected to temperatures normally encountered in Central European aquatic systems. The influence on growth, morphology, membrane fluidity, proteins, and metabolism was examined.     

Intracellular parasitism,the driving force of evolution of Legionella pneumophila and the genus Legionella

Legionella pneumophila is an intracellular pathogen that causes a severe pneumonia called Legionnaires' disease that is often fatal when not promptly diagnosed and treated. Legionella parasitize aquatic protozoa with which it co-evolved over an evolutionary long time. The close relationship between hosts and pathogens, their co-evolution, led to molecular interactions such as the exchange of genetic material through horizontal gene transfer (HGT). Genome sequencing of L. pneumophila and of the entire genus Legionella that comprises over 60 species revealed that Legionellae have co-opted genes and thus cellular functions from their eukaryotic hosts to a surprisingly high extent. Acquisition and loss of these eukaryotic-like genes and domains is an on-going process underlining the highly dynamic nature of the Legionella genomes. Although the large amount and diversity of HGT in Legionella seems to be unique in the prokaryotic world the analyses of more and more genomes from environmental organisms and symbionts of amoeba revealed that such genetic exchanges occur among all amoeba associated bacteria and also among the different microorganisms that infect amoeba. This dynamic reshuffling and gene-acquisition has led to the emergence of Legionella as human pathogen and may lead to the emergence of new human pathogens from the environment.     

Differentiation of Pathogenic and Saprophytic Leptospires I. Growth at Low Temperatures

The minimal growth temperature of the pathogenic leptospires is between 13 and 15 C. The saprophytic leptospires have a minimal growth temperature between 5 and 10 C, or approximately 5 C below that of the pathogens. The capability of the saprophytic leptospires to grow at temperatures below those which allow the growth of the pathogenic leptospires provides a simple method of discrimination. With an inoculum yielding approximately 8 x 10(7) cells per ml in the test medium and an incubation temperature of 13 C, the saprophytic leptospires were easily differentiated from the pathogenic leptospires. All 13 saprophytic leptospires tested grew in the 10% rabbit serum medium at 13 C, whereas none of the 20 pathogens grew during the 30-day incubation period.     

The Tomato Fruit: Import, Growth, Respiration and Carbon Metabolism at Different Fruit Sizes and Temperatures

Measurements of a complete carbon balance sheet over a 48 hperiod for growing tomato fruits at different fruit sizes andtemperature have been carried out. The rates of carbon import,respiration, and growth have been calculated and related toeach other and to the levels of certain carbon metabolites inthe fruit. It was found that there is an excellent linear relationshipbetween the import rate and the sucrose level in the fruit,consistent with the hypothesis that, for the tomato fruit, carbonflows down the sucrose concentration gradient at a rate proportionalto the gradient. This agrees with the findings of Mason andMaskell in cotton. Moreover, the resistance to transport wasrelatively independent of fruit size and temperature. The usualanalysis of respiration in terms of growth and maintenance componentsallowed the determination of conversion efficiencies and maintenancecoefficients for different fruit sizes and temperatures. Asobserved by other authors with other plants, the growth conversionefficiencies were temperature-independent, whereas the maintenancecoefficients were strongly temperature-dependent. The overallconversion efficiency was optimum at 25°C. The specificgrowth rate and the starch level in the tomato fruit were foundto be related.     

非编码RNA与肝脏糖脂代谢调控

随着经济的迅速发展、人们饮食习惯的改变和身体活动的减少,糖尿病成为了现代社会的非传染性的流行病,给家庭和社会造成了极大的危害和经济负担。其中以全身性胰岛素抵抗及胰岛功能衰竭为主要发病特征的2型糖尿病(type 2 diabetes,T2D)已在世界范围引起广泛关注。T2D的发生发展涉及许多组织及糖代谢的各个环节,遗传因素和环境因素共同引起的糖脂代谢通路任一环节的失调均可导致T2D的发生。近年来,包括microRNA(miRNA)及长非编码RNA(LncRNA)在内的非编码RNA(ncRNA)的发现及其在人体生理和病理生理过程中的重要调控作用不断被揭示,为进一步了解T2D的发病机制注入了新理念和信息。miRNAs及LncRNAs的表达具有组织特异性,其表达水平的异常通常与疾病相关。本文主要对miRNAs和LncRNAs在肝脏糖脂代谢调控及T2D的发生发展中的作用及机制的最新研究进展作简要综述。     

A Freeze-Fracture Study of the Cell Membranes of Wheat Adapted to Extracellular Freezing and to Growth at Low Temperatures

Seedlings of Triticum aestivum L. cv. Lennox were grown in natural(late autumn) or controlled environments differing in temperature,to give plants differing either only in growth adaptation oralso in hardiness. The frost hardiness of laminae and leaf baseswas measured using an ion leakage method. Pieces of laminaeand leaf bases were freeze-fixed without chemical cryo-protectantor chemical fixative. The number of particles per unit area of the E fracture faceof the plasma membrane was substantially reduced in laminaefrom the environment inducing most hardening, compared to unhardenedcontrols. This difference in frequency of particles on the Eface was not related to adaptation of growth to low temperature(in the leaf bases) and was not an artificial consequence ofchemical treatment prior to freeze-fixation. The density ofparticles on the P face of the plasma membrane was unaffectedby growth environment. More limited data suggested the densityof particles on either fracture face of the thylakoids was alsounaffected. Unlike an earlier report, particle-free areas ofplasma membrane were not found in hardened tissue. Environment did not affect (a) the width of the ‘mouth’at the plasma membrane-plasmodesma junction in the leaf basesor (b) the size of the nuclear pores. Membrane accumulated belowthe plasma membrane only in some cells from the hardiest material. Key words: Hardiness, Membranes, Triticum     

Impact of Chlorine and Heat on the Survival of Hartmannella vermiformis and Subsequent Growth of Legionella pneumophila

Hartmannella vermiformis, a common amoebal inhabitant of potable-water systems, supports intracellular multiplication of Legionella pneumophila and is probably important in the transportation and amplification of legionellae within these systems. To provide a practical guide for decontamination of potable-water systems, we assessed the chlorine and heat resistance of H. vermiformis. H. vermiformis cysts and trophozoites were treated independently with chlorine at concentrations of 2.0 to 10.0 ppm for 30 min and then cocultured with L. pneumophila. Both cysts and trophozoites were sensitive to concentrations between 2.0 and 4.0 ppm and above (trophozoites somewhat more so than cysts), and 10.0 ppm was lethal to both forms. Hartmannellae treated with chlorine up to a concentration of 4.0 ppm supported the growth of legionellae. To determine whether heat would be an effective addendum to chlorine treatment of amoebae, hartmannellae were subjected to temperatures of 55 and 60°C for 30 min and alternatively to 50°C followed by treatment with chlorine at a concentration of 2 ppm. Fewer than 0.05% of the amoebae survived treatment at 55°C, and there were no survivors at 60°C. Pretreatment at 50°C appeared to make hartmannella cysts more susceptible to chlorine but did not further reduce the concentration of trophozoites.     

Differential Involvement of the Five RNA Helicases in Adaptation of Bacillus cereus ATCC 14579 to Low Growth Temperatures

Bacillus cereus ATCC 14579 possesses five RNA helicase-encoding genes overexpressed under cold growth conditions. Out of the five corresponding mutants, only the ΔcshA, ΔcshB, and ΔcshC strains were cold sensitive. Growth of the ΔcshA strain was also reduced at 30°C but not at 37°C. The cold phenotype was restored with the cshA gene for the ΔcshA strain and partially for the ΔcshB strain but not for the ΔcshC strain, suggesting different functions at low temperature.Bacillus cereus is a human pathogenic sporulated bacterium which is associated with emetic and diarrheal types of food-borne illnesses (4). B. cereus is widespread in the environment and in a wide range of foods. The growth domains of B. cereus strains range from psychrotrophic to nearly thermophilic and correlate with several phylogenetic clusters (15), which presumably permit B. cereus to colonize many different habitats with different thermal regimes. Many foods are stored refrigerated before consumption, and in such cases, B. cereus has to adapt to low-temperature conditions.B. cereus growth at low temperature takes place with a lag phase which may correspond to an adaptation phase (12). Cold is a stress which dramatically affects membrane fluidity, protein synthesis, and also the topology of nucleic acids (22). When exposed to low temperature, bacteria have to face a transient inhibition of protein synthesis mainly due to the presence of secondary structures in mRNA that are stabilized by cold conditions (16, 19). To overcome the translation interruption, cold-shocked cells synthesize cold-induced RNA helicases, which remove secondary structures from RNA duplexes in the presence of ATP, such as CsdA of Escherichia coli (19) or CshA of Bacillus subtilis (1). csdA and srmB deletion mutants of E. coli showed a cold-sensitive phenotype, and these RNA helicases have been described as involved in the biogenesis of the ribosomal 50S subunit at 20°C (10, 11). RNA helicases could also be involved in the degradation of mRNA by unwinding double-stranded mRNA, thereby allowing the action of RNase (8).We have recently shown that the deregulation of the expression of one RNA helicase gene of B. cereus ATCC 14579 increased the lag phase of B. cereus at a low temperature (7). In this context, our aim was to investigate the role of the five putative RNA helicases present in the genome of B. cereus ATCC 14579 in its adaptation at low temperature, close to the growth limit.     

Intermediary Metabolism and Life History Trade-offs: Lipid Metabolism in Lines of the Wing-polymorphic Cricket, Gryllus firmus, Selected for Flight Capability vs. Early Age Reproduction

The extent to which modifications in intermediary metabolismcontribute to life history variation and trade-offs is an importantbut poorly understood aspect of life history evolution. Artificialselection was used to produce replicate genetic stocks of thewing-polymorphic cricket, Gryllus firmus, that were nearly pure-breedingfor either the flight-capable (LW[f]) morph, which delays ovariangrowth, or the flightless (SW) morph, which exhibits enhancedearly-age fecundity. LW(f) lines accumulated substantially moretriglyceride, the main flight fuel in Gryllus, compared withSW-selected lines, and enhanced accumulation of triglyceridewas strongly associated with reduced ovarian growth. Increasedtriglyceride accumulation in LW(f) lines resulted from elevatedde novo biosynthesis of fatty acid and two morph-specific trade-offs:(1) greater proportional utilization of fatty acid for glyceridebiosynthesis vs. oxidation, and (2) a greater diversion of fattyacids into triglyceride vs. phospholipid biosynthesis. Eventhough SW lines produced less total lipid and triglyceride,they produced more phospholipid (important in egg development)than did LW(f) lines. Differences between LW(f) and SW morphsin lipid biosynthesis resulted from substantial alterationsin the activities of all studied lipogenic enzymes, a resultthat is consistent with expectations of Metabolic Control Theory.Finally, application of a juvenile hormone analogue to LW(f)females produced a striking SW phenocopy with respect to allaspects of lipid metabolism studied. Global alterations of lipidmetabolism, most likely produced by alterations in endocrineregulation, underlie morph specializations for flight vs. early-agefecundity in G. firmus. Modification of the endocrine controlof intermediary metabolism is likely to be an important mechanismby which intermediary metabolism evolves and contributes tolife history evolution.