Strict co-linearity of genetic and protein folding domains in an intragenically duplicated rat lens γ-crystallin gene

Two complementary DNA clones pRLγ-2 and pRLγ-3 of different rat lens γ-crystallin messenger RNAs have been used to identify γ-crystallin gene sequences in rat genomic DNA. Subsequently, the DNA present in the 18,000 to 20,000 bases region of the EcoRI digest, giving rise to a strong doublet hybridization signal, was cloned in λ phage Charon-4A. One of the clones, λRCHγ-3, carrying an insert of 17,500 bases has been characterized in detail. From analysis at the restriction enzyme level with 5′-, “middle” and 3′-specific subprobes of pRLγ-3 it could be deduced that λRCHγ-3 contains only one γ-crystallin gene. The coding sequences of this gene are interrupted by intronic DNA. The primary structure of this gene and its flanking regions have been established by sequencing the relevant regions of a subclone of λRCHγ-3, designated pRCHγ-3.1. The sequence data show that the γ-crystallin gene extends over 2700 bases of rat genomic DNA. The gene is split by two introns, one of 87 base-pairs after the third translation codon and a large one of 1880 base-pairs after codon 84. The mosaic structure of the gene is strictly co-linear with the structure of the γ-crystallin polypeptide in that the large intron is positioned in a region which specifies the so-called “connecting peptide” and which links the two highly symmetrical and homologous protein domains. Although expected from the cDNA and protein sequence no introns were observed between the coding regions in the DNA specifying the two homologous folding motifs present in each protein domain. The relevance of this phenomenon in terms of the evolution of the mature γ-crystallin gene is discussed.     

Inactivation of Thirty Viruses by Gamma Radiation

Decimal reduction values (D value) for 30 viruses were determined. The weighted D values of the viruses suspended in Eagle's minimum essential medium ranged from 0.39 to 0.53 Mrads. It was necessary to increase the radiation dose by a factor of >3 to inactivate virus suspended in Eagle's minimum essential medium as compared to the same virus suspended in distilled water. The destruction rate curves were of a first-order reaction.     

Comparative Effect of Temperature on the Oxidative Metabolism of Whole and Disrupted Cells of a Psychrophilic and a Mesophilic Species of Bacillus

We investigated the influence of temperature, in the range of 45 to 5 C, on the rate of oxidation of glucose and citrate by intact cells and cell-free extracts of psychrophilic Bacillus psychrophilus and mesophilic B. thuringiensis. Both glucose and endogenous oxidation by whole and disrupted cells of the psychrophile decreased more slowly with decrease in temperature than did glucose and endogenous oxidation by whole and disrupted cells of the mesophile. Similar results were obtained for the oxidation of citrate by cell-free extracts. Since substrate permeability is not involved in the oxidative metabolism of the cell-free extracts, we concluded that the internal enzymes of the psychrophile differ from those of the mesophile.     

Prokaryotic and eukaryotic pyridoxal-dependent decarboxylases are homologous

Summary A database search has revealed significant and extensive sequence similarities among prokaryotic and eukaryotic pyridoxal phosphate (PLP)-dependent decarboxylases, includingDrosophila glutamic acid decarboxylase (GAD) and bacterial histidine decarboxylase (HDC). Based on these findings, the sequences of seven PLP-dependent decarboxylases from five different organisms have been aligned to derive a consensus sequence for this family of enzymes. In addition, quantitative methods have been employed to calculate the relative evolutionary distances between pairs of the decarboxylases comprising this family. The multiple sequence analysis together with the quantitative results strongly suggest an ancient and common origin for all PLP-dependent decarboxylases. This analysis also indicates that prokaryotic and eukaryotic HDC activities evolved independently. Finally, a sensitive search algorithm (PROFILE) was unable to detect additional members of this decarboxylase family in protein sequence databases.     

The effect of feedback-assisted reduction in heart rate reactivity on videogame performance

In 67 male volunteers, we examined the reduction of cardiovascular responsivity to a psychomotor challenge (videogame) achieved by use of heart rate (HR) feedback and effects of these procedures on concomitant behavioral performance. Each subject participated in a pretraining assessment of his cardiovascular responses to the videogame, a training condition, and a posttraining assessment identical to the initial evaluation. During training, subjects were assigned to one of four conditions: (a) a habituation control group receiving no instructions to alter HR (HC); (b) an instructions-only control group receiving instructions to maintain a low or unchanged HR during videogame presentations (IC); (c) a feedback group receiving instructions to reduce HR using ongoing HR feedback (FB–); or (d) a feedback group receiving instructions to lower HR and given HR feedback plus a score contingency in which total game score was jointly determined by subjects' game performance and success at HR control (FB+). Subjects receiving feedback (FB+, FB–) exhibited greater reductions in HR response to the videogame in the posttraining assessment than control (HC, IC) subjects; FB+ subjects showed greater HR reductions than subjects in any other group. FB+ and FB– subjects showed a lower SBP at posttraining relative to the two control groups, but no reduction in task-induced blood pressure reactivity. There were no group differences in videogame performance, either before or following training.The authors wish to thank Fred Claus, who served as a research assistant for this study.     

Activation of recA protein: The salt-induced structural transition

Purified recA protein is induced by high salt concentrations to hydrolyse ATP even in the absence of DNA. By small angle neutron scattering we show that this salt activation results from a structural transition of the protein filament in the presence of ATPγS from the inactive, compact form (a helical polymer of pitch 70 Å and cross-sectional radius of gyration R_c 40 Å) to the open form (a helical filament of pitch 95 Å and R_c 35 Å, which are the same structural parameters as in the ATPase active complex with DNA and ATP), without detectable change in the degree of association. We conclude that activation of recA is due to the same structural change whether induced by the binding of DNA or by salt. Indeed, the other enzymatic activity of recA, the proteolytic cleavage of the lexA repressor, is found to be inducible by the same salt concentrations as those of the structural transition.     

Effect of a biofilm on the adsorption of 4-chlorophenol on activated carbon

  The adsorption of 4-chlorophenol (4-CP) on activated carbon was studied experimentally both in the presence and in the absence of an inactivated anaerobic biofilm on the surface of carbon pellets. The presence of the biofilm markedly decreased the rate of 4-CP adsorption. However, the final near-equilibrium state (at 27 h) was not affected, and the incremental amount of material adsorbed on the pellets was similar both in the presence and in the absence of the biofilm. The biosorption of 4-CP by a biofilm coating non-adsorbing pellets was also determined. It appears that the biofilm also has some adsorption capability. Freundlich-type equations were used to correlate all data, and transient and near-equilibrium isotherms were obtained for 4-CP adsorption on different adsorbing materials at different times. Received: 30 April 1996 / Received revision: 17 July 1996 / Accepted: 4 August 1996     

Genetically stable cell lines of cucumber for the large-scale production of diploid somatic embryos

For the initiation of embryogenic cucumber ( Cucumis sativus L.) cell lines, from excised radicles, directly in liquid medium, the culture regime, explant density and type and concentration of hormones were adjusted so that pro-embryogenic masses (PEMs) were formed within about 8 weeks. The established cucumber cell lines were maintained tor several years without loss of embryogenic and genetic stability. The ploidy level of somatic embryos from different cucumber eell lines was either diploid or tetraploid and depended on the ploidy level of Ihe cell line. Cucumber cell lines that produced only diploid embryos were obtained by selecting completely diploid explant material and growing it in the dark during the initiation phase. Mixoploid explains could lead to tetraploid or mixoploid ceil lines. Isolation and additional selection and subculturing of single PEMs resulted in either completely diploid or tetraploid cell lines, indicating that all cells of individual PEMs are either diploid or tetraploid. The ernbryogenic cucumber cell Imes, differing only in ploidy level, were indistinguishable in growth rate and embryogetiic potential and were genetically stable over several years.