The amino acid sequence of a Bacillus subtilis phosphoprotein that matches an orfY-tsr coding sequence

Bacillus subtilis contains a 30 kDa protein which was phosphorylated during late vegetative growth and sporulation. The sequence for the N-terminal 16 amino acids was found to be identical to the predicted sequence for the N-terminus of a small open reading frame, orfY, but diverged from the predicted sequence thereafter. The orfY region was resequenced and contained one less adenine residue than previously reported, resulting in an open reading frame from within orfY through the entire coding region for tsr which follows orfY. The predicted orfY-tsr amino acid sequence showed 24% identity to Escherichia coli fructose-1,6-bisphosphate aldolase. Two mutants in the tsr region had 2-5% of wild-type aldolase and the nucleotide sequences showed missense mutations. These results indicate that orfY-tsr encodes aldolase and should be renamed fba1.     

Ingestion,Digestion and Assimilation Efficiency of the Sea-Anemone Aiptasia pallida Fed Zooplankton Prey

Ingestion time, digestion time, and assimilation efficiency by the sea anemone Aiptasia pallida were studied in the laboratory by feeding individual anemones preweighed pellets of freeze-dried Artemia salina nauplii. There was no significant correlation between anemone size, measured as dry weight, and either ingestion time, digestion time or assimilation efficiency. Similarly, there was no significant correlation between meal size (i.e., dry weight of ingested brine shrimp pellet) and either ingestion time, digestion time or assimilation efficiency. These results suggest that, under these conditions, assimilation efficiency is unaffected by either “meal” size or anemone size.     

Differences in neuroinvasion and protective innate immune pathways between encephalitic California Serogroup orthobunyaviruses

The California serogroup (CSG) of Orthobunyaviruses comprises several members capable of causing neuroinvasive disease in humans, including La Crosse orthobunyavirus (LACV), Jamestown Canyon orthobunyavirus (JCV), and Inkoo orthobunyavirus (INKV). Despite being genetically and serologically closely related, their disease incidences and pathogenesis in humans and mice differ. We have previously shown that following intraperitoneal inoculation of weanling mice, LACV was highly pathogenic while JCV and INKV were not. To determine why there were differences, we examined the ability of these viruses to invade the CNS and compared the host innate immune responses that regulated viral pathogenesis. We found that LACV was always neuroinvasive, which correlated with its high level of neuroinvasive disease. Interestingly, JCV was not neuroinvasive in any mice, while INKV was neuroinvasive in most mice. The type I interferon (IFN) response was critical for protecting mice from both JCV and INKV disease, although in the periphery JCV induced little IFN expression, while INKV induced high IFN expression. Despite their differing neuroinvasive abilities, JCV and INKV shared innate signaling components required for protection. The presence of either cytoplasmic Rig-I-Like Receptor signaling or endosomal Toll-Like Receptor signaling was sufficient to protect mice from JCV or INKV, however, inhibition of both pathways rendered mice highly susceptible to neurological disease. Comparison of IFN and IFN-stimulated gene (ISG) responses to INKV in the brains of resistant wild type (WT) mice and susceptible immune knockout mice showed similar IFN responses in the brain, but WT mice had higher ISG responses, suggesting induction of key ISGs in the brain is critical for protection of mice from INKV. Overall, these results show that the CSG viruses differ in neuroinvasiveness, which can be independent from their neuropathogenicity. The type I IFN response was crucial for protecting mice from CSG virus-induced neurological disease, however, the exact correlates of protection appear to vary between CSG viruses.     

Effects of Altered Light and Feeding Regimes on the Zooplankton Feeding Capability of Hydra viridis

The effects of light regime, feeding regime and tentacle number on the zooplankton feeding capability of Hydra viridis were tested in the laboratory. Feeding was measured by exposing Hydra to a known volume of Artemia salina nauplii and recording the number captured and ingested. In all cases there was a correlation between the number of Artemia captured and the number ingested. H. viridis with 7 tentacles captured and ingested more Artemia than Hydra with 6 tentacles. However, changes in light and/or feeding regimes did not alter the number of tentacles/Hydra. Varying light and feeding regimes altered the number of Zoochlorellae/cell and Hydra growth rate. There was no effect on the number of Artemia captured or ingested and no effect on the percent ingestion of captured Artemia. These data suggest that, under these conditions, zooplankton feeding by H. viridis is independent of nutritional history.     

Relations between pigments and proteins in the photosynthetic membranes of Rhodopseudomonas spheroides

We have isolated from Rhodopseudomonas spheroides a pigment-protein complex of apparent weight 9 kdaltons that bears more than 60% of the light harvesting bacteriochlorophyll. The isolation procedure involved exposure to 1% lauryl dimethyl amine oxide (LDAO). The purified 9-kdalton fraction showed the light harvesting bacteriochlorophyll components B800 and B850, plus carotenoids. The ratio of bacteriochlorophyll to protein was 17%. This protein is probably the same as the “band 15” protein of Fraker and Kaplan. It may exist in vivo as characteristic aggregates of higher molecular weight. LDAO added to Rps. spheroides chromatophores converted the bacteriochlorophyll component B870 to a form absorbing at 770 nm but had little effect on the “B800 + B850” system, causing only a reversible shift of the 850-nm band to 845 nm. Anti-reaction center serum, added to subcellular fractions from Rps. spheroides with 1% LDAO, precipitated reaction center chromoprotein unaccompanied by light harvesting bacteriocholorophyll. Other antisera precipitated light harvesting components and left the reaction center chromophores in solution. A major protein of apparent weight 45 kdaltons was found in relatively nonpigmented fractions from Rps. spheroides, associated with cell wall fragments. The 45-kdalton protein showed considerable interstrain variability, whereas the 9-kdalton and reaction center proteins appeared constant.     

Motivational control of caching behaviour in the scrub jay, Aphelocoma coerulescens

We investigated the motivational control of caching behaviour in scrub jays using a two-stage procedure to examine the effects of prefeeding and/or precaching (stage 1) on subsequent caching behaviour (stage 2). Experiment 1 demonstrated that both prefeeding and precaching reduced the subsequent caching of both edible (peanuts) and inedible (stones) items. The reduction in caching was greatest when the items available for storing were the same in the two stages. This item specificity was confirmed in experiment 2 using two food types, peanuts and dog food kibbles. The final experiment demonstrated that the effect of prefeeding on subsequent caching can also be food specific, in that birds that received food in a powdered form that they could eat, but not cache in stage 1, showed a reduction in subsequent caching in stage 2 only when the food type was the same in the two stages. These results suggest that caching behaviour is controlled by both the feeding system and an independent caching system, and that this control is mediated by the incentive value of the specific items rather than by a general motivational state. Copyright 1999 The Association for the Study of Animal Behaviour.     

Genomic instability and enhanced radiosensitivity in Hsp70.1- and Hsp70.3-deficient mice

Heat shock proteins (HSPs) are highly conserved among all organisms from prokaryotes to eukaryotes. In mice, the HSP genes Hsp70.1 and Hsp70.3 are induced by both endogenous and exogenous stressors, such as heat and toxicants. In order to determine whether such proteins specifically influence genomic instability, mice deficient for Hsp70.1 and Hsp70.3 (Hsp70.1/3(-/-) mice) were generated by gene targeting. Mouse embryonic fibroblasts (MEFs) prepared from Hsp70.1/3(-/-) mice did not synthesize Hsp70.1 or Hsp70.3 after heat-induced stress. While the Hsp70.1/3(-/-) mutant mice were fertile, their cells displayed genomic instability that was enhanced by heat treatment. Cells from Hsp70.1/3(-/-) mice also display a higher frequency of chromosome end-to-end associations than do control Hsp70.1/3(+/+) cells. To determine whether observed genomic instability was related to defective chromosome repair, Hsp70.1/3(-/-) and Hsp70.1/3(+/+) fibroblasts were treated with ionizing radiation (IR) alone or heat and IR. Exposure to IR led to more residual chromosome aberrations, radioresistant DNA synthesis (a hallmark of genomic instability), increased cell killing, and enhanced IR-induced oncogenic transformation in Hsp70.1/3(-/-) cells. Heat treatment prior to IR exposure enhanced cell killing, S-phase-specific chromosome damage, and the frequency of transformants in Hsp70.1/3(-/-) cells in comparison to Hsp70.1/3(+/+) cells. Both in vivo and in vitro studies demonstrate for the first time that Hsp70.1 and Hsp70.3 have an essential role in maintaining genomic stability under stress conditions.