Onset of deaminase APOBEC3B induction in response to DNA double-strand breaks

Deamination of 5-methyl cytosine is a major cause of cancer-driver mutations in inflammation-associated cancers. The deaminase APOBEC3B is expressed in these cancers and causes mutations under replication stress; however, the mechanisms by which APOBEC3B mediates deamination and its association with genomic disorders are still unclear. Here, we show that APOBEC3B is stabilized to induce deamination reaction in response to DNA double-strand breaks (DSBs), resulting in the formation of long-lasting DSBs. Uracil, the major deamination product, is subsequently targeted by base excision repair (BER) through uracil-DNA glycosylase 2 (UNG2); hence late-onset DSBs arise as by-products of BER. The frequency of these delayed DSBs was increased by treatment of cells with a PARP inhibitor, and was suppressed following knock-down of UNG2. The late-onset DSBs were induced in an ATR-dependent manner. Those secondary DSBs were persistent, unlike DSBs directly caused by γ-ray irradiation. Overall, these results suggest that the deaminase APOBEC3B is induced in response to DSBs, leading to long-lasting DSB formation in addition to mutagenic 5me-C>T transition induction.     

Nitrogen-stable isotopic signatures of basal food items, primary consumers and omnivores in rivers with different levels of human impact

We examined how nitrogen-stable isotopic signatures of food web components (basal resources, primary and lower consumers, and omnivores) in rivers change with increasing levels of human population density (HPD) in their watersheds. Samples were collected from 22 rivers flowing in the Lake Biwa basin, Japan. Among three potential resources at the base of food webs (epilithon, benthic and suspended particulate organic matter), the mean isotopic values (δ¹⁵N) of the epilithon (4.5–7.8%) were consistently higher than those of other items (1.9–4.2%) and displayed the most pronounced elevation (by 3.3%) with increasing HPD. The mean δ¹⁵N values of the individual taxa of lower consumers (bivalve, snail and caddisfly) tended to increase with increasing HPD, although the pattern and the extent of the elevation were highly variable among the taxa. These results suggest a taxon-specific feature in the N source (or sources) of lower consumers. Our data suggested that human activities (e.g. nutrient loading) potentially induce changes in the N baselines of river food webs. The major N source of bivalves appeared to be shifted from suspended particulate organic matter to other items with increasing HPD. Trophic levels of goby fish (Rhinogobius sp. OR) and shrimp (Palaemon paucidens), being estimated to be at 2.4–3.8 and 2.1–3.4, respectively, did not differ significantly among rivers with different HPD levels. An erratum to this article can be found at     

GAPDH-A Recruits a Plant Virus Movement Protein to Cortical Virus Replication Complexes to Facilitate Viral Cell-to-Cell Movement

The formation of virus movement protein (MP)-containing punctate structures on the cortical endoplasmic reticulum is required for efficient intercellular movement of Red clover necrotic mosaic virus (RCNMV), a bipartite positive-strand RNA plant virus. We found that these cortical punctate structures constitute a viral replication complex (VRC) in addition to the previously reported aggregate structures that formed adjacent to the nucleus. We identified host proteins that interacted with RCNMV MP in virus-infected Nicotiana benthamiana leaves using a tandem affinity purification method followed by mass spectrometry. One of these host proteins was glyceraldehyde 3-phosphate dehydrogenase-A (NbGAPDH-A), which is a component of the Calvin-Benson cycle in chloroplasts. Virus-induced gene silencing of NbGAPDH-A reduced RCNMV multiplication in the inoculated leaves, but not in the single cells, thereby suggesting that GAPDH-A plays a positive role in cell-to-cell movement of RCNMV. The fusion protein of NbGAPDH-A and green fluorescent protein localized exclusively to the chloroplasts. In the presence of RCNMV RNA1, however, the protein localized to the cortical VRC as well as the chloroplasts. Bimolecular fluorescence complementation assay and GST pulldown assay confirmed in vivo and in vitro interactions, respectively, between the MP and NbGAPDH-A. Furthermore, gene silencing of NbGAPDH-A inhibited MP localization to the cortical VRC. We discuss the possible roles of NbGAPDH-A in the RCNMV movement process.     

Changes in expression of provasotocin and proisotocin genes during adaptation to hyper- and hypo-osmotic environments in rainbow trout

Summary The physiological roles of neurohypophysial hormones, vasotocin (VT) and isotocin (IT), are not yet clear in teleosts. Since information on responsiveness of hypothalamic neurosecretory neurons to environmental stimuli may contribute to an understanding of their physiological roles, effects of environmental hyper- and hypo-osmotic stimuli on expression of VT and IT precursor (proVT and proIT) genes in rainbow trout were investigated, using an in situ hybridization technique in which 46 mer synthetic oligonucleotides were used as hybridization probes. The probes corresponded to the mRNA loci encoding chum salmon proVT (-5 to 11) and proIT (-5 to 11), and were labeled at the 3-end with ³⁵S. Autoradiographic silver grains which represent the hybridization signals of proVT and proIT mRNAs were localized in both magnocellular and parvocellular neurons in the nucleus preopticus magnocellularis (NPOmg). Localizations of proVT and proIT hybridization signals coincided with those of VT- and IT-immunoreactive neurons in adjacent sections, and showed that proVT and proIT genes are expressed in separate neurons. The intensity of proVT hybridization signals as determined by grain counting in magnocellular neurons in the NPOmg was conspicuously decreased after transfer from fresh water (FW) to 80% seawater (SW). The proVT mRNA levels in SW trout were consistently lower than those of FW trout for up to 2 weeks. After return from 80% SW to FW, the proVT mRNA level increased, attaining the initial FW level. The proIT mRNA levels in SW trout were not statistically different from those in FW trout, except for the 1st day after transfer to SW. These results suggest that synthesis of proVT was elevated by transfer from higher to lower salinity, and that VT may have a physiological role in salmonid osmoregulation, especially in adaptation to a hypo-osmotic environment.Abbreviations ABC avidin-biotin-peroxidase complex - AVP arginine vasopressin - BSA bovine serum albumin - EDTA ethylene diamine tetra-acetic acid - cpm counts per minute - FW fresh water - GFR gromerular filtration rate - ISH in situ hybridization - IT isotocin - mRNA messenger ribonucleic acid - NPOmg nucleus preopticus magnocellularis - OXT oxytocin - PBS phosphate-buffered saline - SSC saline sodium citrate - RT room temperature - SW seawater - VT vasotocin     

Effects of Volatiles Evolved from Excised Plant Tissues and Lemon Oil on the Growth of Mung Bean Seedlings

Volatiles evolved from excised segments of leaves of oleander(Nerium oleander) and cedar (Cedrus deodara), peels of lemon(Citrus limon), orange (Citrus sinensis) and lime (Citrus aurantifolia),and roots of wasabi (Wasabia japonica) inhibited growth of mungbean seedlings. The volatiles evolved from lemon oil also markedlyinhibited elongation of mung bean seedlings. The active constituent(s)in the volatiles were distinct from ethylene. The mode of inhibitionby the volatiles from lemon oil was different from that of ethylene.The volatiles from lemon oil alleviated the inhibitory effectof ethylene on the growth of seedlings. The volatiles from lemonoil supressed the formation of lamellar structure in chloroplasts,thus resulting in etiolation of seedlings. (Received October 25, 1982; Accepted December 29, 1982)     

Biochemical Effects of Gamma Radiation on Potato and Sweet Potato Tissues

Relationship between the chemical constituents of tobacco leaves and the gaseous constituents of cigarette smoke from which K value¹⁾ was computed was discussed and the following presuppositions were demonstrated to be correct.

1. Fibrous substances in tobacco leaves are the main precursors of acetaldehyde, propionaldehyde, acrolein, acetone, methylethylketone, diacetyl, methanol, furan, an unknown compound, No. 6 and an unknown compound, No. 16 in cigarette smoke.

2. Sugars in tobacco leaves are the main precursors of 2-methylfuran and 2,5-dimethyl- furan in cigarette smoke.

3. Resinous substances in tobacco leaves are the main precursors of isoprene and an unknown compound, No. 2 in cigarette smoke.

     

Wound-Induced Ethylene Production and 1-Aminocyclopropane-1-carboxylic Acid Synthase in Mesocarp Tissue of Winter Squash Fruit

Discs (9 mm in diameter and 2 mm in thickness) sliced from mesocarpof winter squash fruit (Cucurbita maxima Duch.) upon incubationat 24°C produced ethylene at an increasing rate after alag period of 3 h. 1-Aminocydopropane-l-carboxylic acid (ACC)synthase activity also increased at a rapid rate after lag periodof less than 3 h, reaching a peak 14 h after incubation andthen declining sharply. The rise in ACC synthase activity precededa rapid increase in ACC formation and ethylene production. Inductionof ACC synthase by wounding in sliced discs was strongly suppressedby the application of cycloheximide, actinomycin D and cordycepin,suggesting that the rise in ACC synthase activity may resultfrom de novo synthesis of protein. ACC synthase extracted from wounded tissue of winter squashmesocarp required pyridoxal phosphate for its maximum activity.The optimum pH of the reaction was 8.5. Km value for S-adenosylmethioninewas 120 µM. The reaction was markedly inhibited by aminoethoxyvinylglycinewith Ki value being 2.7 µM. (Received March 23, 1983; Accepted May 23, 1983)