Defense Responses to Tetrapyrrole-Induced Oxidative Stress in Transgenic Plants with Reduced Uroporphyrinogen Decarboxylase or Coproporphyrinogen Oxidase Activity1

We analyzed the antioxidative defense responses of transgenic tobacco (Nicotiana tabacum) plants expressing antisense RNA for uroporphyrinogen decarboxylase or coproporphyrinogen oxidase. These plants are characterized by necrotic leaf lesions resulting from the accumulation of potentially photosensitizing tetrapyrroles. Compared with control plants, the transformants had increased levels of antioxidant mRNAs, particularly those encoding superoxide dismutase (SOD), catalase, and glutathione peroxidase. These elevated transcript levels correlated with increased activities of cytosolic Cu/Zn-SOD and mitochondrial Mn-SOD. Total catalase activity decreased in the older leaves of the transformants to levels lower than in the wild-type plants, reflecting an enhanced turnover of this photosensitive enzyme. Most of the enzymes of the Halliwell-Asada pathway displayed increased activities in transgenic plants. Despite the elevated enzyme activities, the limited capacity of the antioxidative system was apparent from decreased levels of ascorbate and glutathione, as well as from necrotic leaf lesions and growth retardation. Our data demonstrate the induction of the enzymatic detoxifying defense system in several compartments, suggesting a photosensitization of the entire cell. It is proposed that the tetrapyrroles that initially accumulate in the plastids leak out into other cellular compartments, thereby necessitating the local detoxification of reactive oxygen species.

Intrinsic noise in gene regulatory networks

Cells are intrinsically noisy biochemical reactors: low reactant numbers can lead to significant statistical fluctuations in molecule numbers and reaction rates. Here we use an analytic model to investigate the emergent noise properties of genetic systems. We find for a single gene that noise is essentially determined at the translational level, and that the mean and variance of protein concentration can be independently controlled. The noise strength immediately following single gene induction is almost twice the final steady-state value. We find that fluctuations in the concentrations of a regulatory protein can propagate through a genetic cascade; translational noise control could explain the inefficient translation rates observed for genes encoding such regulatory proteins. For an autoregulatory protein, we demonstrate that negative feedback efficiently decreases system noise. The model can be used to predict the noise characteristics of networks of arbitrary connectivity. The general procedure is further illustrated for an autocatalytic protein and a bistable genetic switch. The analysis of intrinsic noise reveals biological roles of gene network structures and can lead to a deeper understanding of their evolutionary origin.

Genetic mapping of a murine locus controlling development of T helper 1/T helper 2 type responses.

Genetic background of the T cell can influence T helper (Th) phenotype development, with some murine strains (e.g., B10.D2) favoring Th1 development and others (e.g., BALB/c) favoring Th2 development. Recently we found that B10.D2 exhibit an intrinsically greater capacity to maintain interleukin 12 (IL-12) responsiveness under neutral conditions in vitro compared with BALB/c T cells, allowing for prolonged capacity to undergo IL-12-induced Th1 development. To begin identification of the loci controlling this genetic effect, we used a T-cell antigen receptor-transgenic system for in vitro analysis of intercrosses between BALB/c and B10.D2 mice and have identified a locus on murine chromosome 11 that controls the maintenance of IL-12 responsiveness, and therefore the subsequent Th1/Th2 response. This chromosomal region is syntenic with a locus on human chromosome 5q31.1 shown to be associated with elevated serum IgE levels, suggesting that genetic control of Th1/Th2 differentiation in mouse, and of atopy development in humans, may be expressed through similar mechanisms.

Superluminal sources.

Predictions for the apparent velocity statistics under simple beaming models are presented and compared to the observations. The potential applications for tests of unification models and for cosmology (source counts, measurements of the Hubble constant H0 and the deceleration parameter q0) are discussed. First results from a large homogeneous survey are presented. The data do not show compelling evidence for the existence of intrinsically different populations of galaxies, BL Lacertae objects, or quasars. Apparent velocities betaapp in the range 1-5 h-1, where h = H0/100 km.s-1.Mpc-1 [1 megaparsec (Mpc) = 3.09 x 10(22) m], occur with roughly equal frequency; higher values, up to betaapp = 10 h-1, are rather more scarce than appeared to be the case from earlier work, which evidently concentrated on sources that are not representative of the general population. The betaapp distribution suggests that there might be a skewed distribution of Lorentz factors over the sample, with a peak at gammab approximately 2 h-1 and a tail up to at least gammab approximately 10 h-1. There appears to be a clearly rising upper envelope to the betaapp distribution when plotted as a function of observed 5-GHz luminosity; a combination of source counts and the apparent velocity statistics in a larger sample could provide much insight into the properties of radio jet sources.

The case for unification.

I investigate the issue of whether the various subclasses of radio-loud galaxies are intrinsically the same but have been classified differently mainly due to their being viewed from different directions. Evidence for the two key elements of this popular version of the "unified scheme (US)," relativistic jets and nuclear tori, is updated. The case for the torus opening angle increasing with the radio luminosity of the active galactic nucleus (AGN) is freshly argued. Radio-loud AGN are particularly suited for testing the US, since their structures and polarization properties on different scales, as well as their overall radio sizes, provide useful statistical indicators of the relative orientations of their various subclasses. I summarize recent attempts to bring under a single conceptual framework the USs developed for radio-moderate [Fanaroff-Riley type I (FRI)] and radio-powerful (FRII) AGN. By focusing on FRII radio sources, I critically examine the recent claims of conflict with the US, based on the statistics of radio-size measurements for large, presumably orientation-independent, samples with essentially complete optical identifications. Possible ways of reconciling these results, and also the ones based on very-long-baseline radio interferometry polarimetric observations, with the US are pointed out. By incorporating a highly plausible temporal evolution of radio source properties into the US, I outline a scenario that allows the median linear size of quasars to approach, or even exceed, that of radio galaxies, as samples with decreasing radio luminosity are observed. Thus, even though a number of issues remain to be fully resolved, the scope of unified models continues to expand.

The rat suprachiasmatic nucleus is a clock for all seasons.

Seasonal changes of daylength (photoperiod) affect the expression of hormonal and behavioral circadian rhythms in a variety of organisms. In mammals, such effects might reflect photoperiodic changes in the circadian pace-making system [located in the suprachiasmatic nucleus (SCN) of the hypothalamus] that governs these rhythms, but to date no functionally relevant, intrinsic property of the SCN has been shown to be photoperiod dependent. We have analyzed the temporal regulation of light-induced c-fos gene expression in the SCN of rats maintained in long or short photoperiods. Both in situ hybridization and immunohistochemical assays show that the endogenous circadian rhythm of light responsiveness in the SCN is altered by photoperiod, with the duration of the photosensitive subjective night under the short photoperiod 5-6 h longer than under the long photoperiod. Our results provide evidence that a functional property of the SCN is altered by photoperiod and suggest that the nucleus is involved in photoperiodic time measurement.

High-affinity binding sites for histone H1 in plasmid DNA.

The interaction of histone H1 isolated from chicken erythrocytes with restriction fragments from plasmids pBR322 and pUC19 was studied by gel electrophoresis. Certain restriction fragments exhibited unusually high affinity for the histone, forming high molecular mass complexes at protein to DNA ratios at which the other fragments did not show evidence for binding. The highly preferred fragments are intrinsically curved, as judged by their electrophoretic mobility in polyacrylamide gels, by computer modeling, and by imaging with scanning force microscopy. However, control experiments with either curved portions of the same fragments or highly curved kinetoplast DNA fragments showed that the presence of curvature alone was not sufficient for preferential binding. By using various restriction fragments centered around the highly preferred sequence, it was found that the high-affinity binding required in addition the presence of specific sequences on both sides of the region of curvature. Thus, both curvature and the presence of specific sites seem to be required to generate high affinity.