Genomic DNA structure of two new horseradish-peroxidase-encoding genes

Genomic DNAs encoding the horseradish peroxidase (HRP) isozymes, prxC2 and prxC3, were cloned and sequenced. By comparing the sequences of the HRP isozyme-encoding genes, prxC1a and prxC1b and their cDNA [Fujiyama et al., Eur. J. Biochem. 173 (1988) 681-687], , it was concluded that prxC2 and prxC3 consisted of four exons and three introns as in the prxC1 gene family. The position of introns in coding regions were the same in all four prx genes. Genes prxC2 and prxC3 coded for 347 and 349 amino acid (aa) residues, respectively, including putative signal sequences at the N termini. In the flanking regions of both genes, putative promoters and polyadenylation signals were found. Nucleotide sequence homology in the coding region was 71% between prxC1a and prxC2, and 66% between prxC1a and prxC3. The aa sequence homologies in plant and microbial peroxidases were compared.     

The nylon oligomer biodegradation system ofFlavobacterium andPseudomonas

This review article is a compendium of the available information on the degradation of a man-made compound, 6-aminohexanoate-oligomer, inFlavobacterium andPseudomonas strains, and discusses the molecular basis for adaptation of microorganisms toward these xenobiotic compounds. Three plasmid-encoded enzymes, 6-aminohexanoate-cyclic-dimer hydrolase (EI), 6-aminohexanoate-dimer hydrolase (EII), and endo-type 6-aminohexanoate-oligomer hydrolase (EIII) are responsible for the degradation of the oligomers. Two repeated sequences, designated RS-I and RS-II, are found on plasmid pOAD2, which is involved in 6-aminohexanoate degradation inFlavobacterium. RS-I appears 5 times on the pOAD2, and all copies have the same sequences as insertion sequence IS6100. RS-II appears twice on the plasmid. RS-IIA contains the gene encoding EII, while RS-IIB contains the gene for the analogous EII' protein. Both EII and EII' are polypeptides of 392 amino acids, which differ by 46 amino acid residues. The specific activity of the EII enzyme is 200-fold higher than that of EII'. Construction of various hybrid genes demonstrated that only the combination of two amino acid residues in the EII' enzyme can enhance the activity of the EII' to the same level as that of EII enzyme.Abbreviations EI 6-aminohexanoate-cyclic-dimer hydrolase - EII 6-aminohexanoate-dimer hydrolase - EIII endo-type 6-aminohexanoate-oligomer hydrolase - F-EI EI fromFlavobacterium - F-EII EII fromFlavobacterium - P-EI EI fromPseudomonas - P-EII EII fromPseudomonas - EII' a protein having 88% homology to the EII encoded on the RS-IIB region of pOAD2 - nylA gene for the EI enzyme - nylB gene for the EII enzyme - nylC gene for the EIII enzyme - nylB' gene for the EII' protein - kb kilo-base-pairs     

Premotor-Motor Interhemispheric Inhibition Is Released during Movement Initiation in Older but Not Young Adults

Neural interactions between contralateral motor regions are thought to be instrumental in the successful preparation, and execution, of volitional movements. Here we investigated whether healthy ageing is associated with a change in functional connectivity, as indicated by the ability to modulate interhemispheric interactions during movement preparation in a manner that assists rapid movement responses. Thirteen young (mean age 22.2 years) and thirteen older (68.5 years) adults rapidly abducted their left index finger as soon as possible in response to a visual imperative signal, presented 500 ms after a visual warning signal.Interactions between left dorsal premotor cortex (LPMd) and right primary motor cortex (RM1) and between left primary motor cortex (LM1) and RM1 were investigated at six time points between the warning signal and the volitional response using paired-pulse transcranial magnetic stimulation. Relative to the inhibitory interactions measured at rest, both young and older adults released LM1-RM1 inhibition beginning 250 ms after the warning signal, with no significant differences between groups. LPMd-RM1 interactions became facilitatory (from the onset of the imperative signal onwards) in the older, but not the young, group. Regression analyses revealed that for the older adults, modulation of LPMd-RM1 interactions early in the preparation period was associated with faster responses, suggesting that specifically timed modulation of these pathways may be a compensatory mechanism to offset, at least in part, slowing of motor responses. The results suggest a greater reliance on premotor regions during the preparation of simple motor actions with advancing age.     

Construction of hybrid genes of 6-aminohexanoic acid-oligomer hydrolase and its analogous enzyme. Estimation of the intramolecular regions important for the enzyme evolution

Hybrids were constructed of the genes for two homologous enzymes, 6-aminohexanoic acid-oligomer hydrolase (EII, one of the nylon oligomer degradation enzymes), and its probable evolutionary antecedent (EII'). The structural genes of EII (nylB) and EII' (nylB') have 88% similarity in their nucleotide sequences, an open frame encoding a peptide of 392 amino acids, conserved restriction sites, and in vitro recombination between these genes at the corresponding restriction sites generated genes directing various hybrid enzymes. In a comparison of the EII, EII', and the hybrid enzymes, we concluded that one or more of the four amino acid alterations that occurred in the intramolecular region (between amino acids 162-257) of EII' is essential to the adaptation of the enzyme to nylon oligomer degradation, and that its effect is enhanced 20-fold by one or more further alterations in the 258-380 region. Our results also suggest that this technique is useful for improving enzyme characteristics.     

Expression of human CMP-N-acetylneuraminic acid synthetase and CMP-sialic acid transporter in tobacco suspension-cultured cell

Plant cells have no beta1,4-galactosylated and sialylated glycan, which plays important roles in biological functions in animal cells. Previously, we generated transgenic tobacco BY2 suspension-cultured cells that produced human beta1,4-galactosyltransferase [N.Q. Palacpac, S. Yoshida, H. Sakai, Y. Kimura, K. Fujiyama, T. Yoshida, T. Seki, Stable expression of human beta1,4-galactosyltransferase in plant cells modifies N-linked glycosylation pattern, Proc. Natl. Acad. Sci. USA 96 (1999) 4692-4697]. In this study, we introduced two critical genes encoding human CMP-N-acetylneuraminic acid synthetase and CMP-sialic acid transporter into tobacco suspension-cultured cell to pave a route for sialic biosynthetic pathway. The recombinant human proteins showed their biological activities. These results show that the plant cell can be a useful bioreactor for the production of mammalian glycoproteins.     

Local variation in the thistle species Cirsium grayanum affects its utilization by the herbivorous ladybird beetle Henosepilachna niponica

Host plants are the most critical environmental factor for phytophagous arthropods. Adaptation to a novel host will alter the distributional range of an herbivore to include the area the novel host covers, and might promote divergence between populations utilizing the old and new hosts. On the Oshima Peninsula, Hokkaido Island, northern Japan, the ladybird beetle Henosepilachna niponica (Lewis) (Coleoptera: Coccinellidae) usually occurs exclusively on the thistle Cirsium alpicola Nakai (Asteraceae), which is distributed in the southernmost part of the island, although some other Cirsium species often grow in sympatry. At least at one site (Assabu), however, H. niponica depends on another thistle, Cirsium grayanum (Maxim.) Nakai, the most abundant and widely distributed thistle species on the Oshima Peninsula. We examined adult feeding acceptance and preference, and larval performance, in two populations of H. niponica that utilize different species of thistles (the Assabu population occurs on C. grayanum; the Shiriuchi population, on C. alpicola) by testing three kinds of thistles (C. grayanum from Assabu, C. grayanum from Shiriuchi, and C. alpicola from Shiriuchi). Results strongly suggested that among the three plants tested, C. grayanum from Assabu was the most suitable host for H. niponica. We concluded that local differences in C. grayanum mainly explain why it is not used at Shiriuchi, but that there are also local differences in host use among the beetle populations, suggesting some degree of local adaptation. This suggests that different selective regimes associated with different host plants might promote divergence among beetle populations. We conclude that H. niponica is at present unlikely to expand its range to the whole range of C. grayanum.     

Oviposition site selection by herbivorous beetles: a comparison of two thistle feeders, Cassida rubiginosa and Henosepilachna niponica

In insects that feed on plants in both adult and larval stages, it is often difficult to distinguish oviposition preference from adult feeding preference, because oviposition can occur at or in proximity to feeding sites. In the present study, characteristics of oviposition site selection of two beetle species, Cassida rubiginosa Müller (Coleoptera: Chrysomelidae) and Henosepilachna niponica (Lewis) (Coleoptera: Coccinellidae), were investigated in the field and laboratory, with particular attention to relationships with adult feeding sites. In the field, distances between adult feeding scars and egg masses differed for C . rubiginosa and H . niponica , with the former being very small and the latter averaging 24.6 cm. The same tendencies for the distances between adult feeding scars and egg masses of the two beetle species were confirmed in cages in which only female beetles were released. Cassida rubiginosa restricted egg laying to host plants in the field and to leaves in laboratory assays. On the other hand, H . niponica placed 8% of egg masses on plants adjacent to host plants in the field and often placed eggs on artificial substrates rather than leaf discs in laboratory assays. These results suggest that oviposition and female feeding sites are virtually inseparable in the case of C . rubiginosa , while H . niponica females do not necessarily keep to host plant leaves as oviposition substrates and they tend to oviposit at some distance from their feeding sites. Results are discussed in relation to proximate and ultimate causes of host selection behavior.     

Differentiation in the ability to utilize Pterostyrax hispida (Ebenales: Styracaceae) among four local populations of the phytophagous ladybird beetle Henosepilachna yasutomii (Coleoptera: Coccinellidae)

The oligophagous ladybird beetle Henosepilachna yasutomii Katakura (Coleoptera: Coccinellidae) shows inter-population differences in its host-use. In this beetle, populations using the deciduous tree Pterostyrax hispida Sieb. et Zucc. (Styracaceae) were recently rediscovered in the Kanto districts of central Honshu, Japan. In the present study, the ability to utilize P. hispida and other host plants was compared among four populations of H. yasutomii occurring on P. hispida and Scopolia japonica Maxim. (Solanaceae), Chelidonium japonicum Thunb. (Papaveraceae), and Solanum tuberosum L. (Solanaceae). As regards the feeding habits of adult beetles, only the populations occurring on P. hispida accepted this plants leaves, which differed distinctly from the feeding habits of the other populations, although the differences among the four populations were not readily apparent with respect to the leaf amounts consumed. The larvae from the populations occurring on P. hispida showed significantly higher survivorship on this plant than did the larvae from the other populations. Considering the host use patterns and the life cycles of beetles under natural conditions, the large abundance of P. hispida leaves throughout the season may have played an important role in selection for the ability to utilize P. hispida observed in the H. yasutomii populations occurring on this woody host.     

Distribution of Non-tasters for Phenylthiocarbamide and High Sensitivity to Quinine Hydrochloride of the Non-tasters in Japanese

The percentage of non-tasters for phenylthiocarbamide in 915Japanese students was 9.4%. The thresholds of the edge and backof the tongue to quinine hydrochloride were significantly smallerin the non-tasters than in the tasters. The thresholds of anytongue portions to NaCl, acetic acid or sucrose did not differbetween the tasters and the non-tasters. Chem. Senses 22: 547–551,1997.