A comparative study of spectral sensitivity curves in three diurnal and eight nocturnal species of Japanese fireflies

The spectral properties of the eyes of 3 species of diurnal and 8 species of nocturnal Japanese fireflies, in many cases males and females, were determined by an electroretinographic method. With the exception of Hotaria parvula males, which had a λ_max of 580 nm, almost all species studied possessed a maximum around 500–540 nm. The eyes of diurnal and nocturnal species did not differ significantly in their sensitivity maxima. As in North American species of fireflies (Lall, 1981a,b) congruency existed between visual sensitivity peaks and light emission maxima in Luciola cruciata, L. lateralis and Hotaria parvula. In agreement with Seliger et al. (1982a,b) we conclude that an adaptation of the visual sensitivity to the light produced need not have occurred and that evolutionary adaptation of light emission to an existing ancestral green-sensitivity of the eye is the more likely course of events.     

Two-fold symmetry of crystalline DNA-EcoRI endonuclease recognition complexes

Recognition complexes between EcoRI endonuclease and either of two synthetic oligonucleotides (sequences CGCGAATTCGCG and TCGCGAATTCGCG) crystallize in Space Group P321 with unit cell parameters a = 128 and c = 47 A and a = 118.4 and c = 49.7 A, respectively. Native diffraction data to 3 A resolution have been collected from the form containing the tridecameric sequence. Electrophoretic analyses of dissolved crystals demonstrate that this form contains DNA and protein in a ratio of one double helix per enzyme dimer. The most likely asymmetric unit contents are one 31,000 dalton enzyme subunit and one strand of DNA, yielding VM values of 3.1 A3/dal and 2.8 A3/dal for the forms containing dodecameric and tridecameric DNA, respectively. This implies that the DNA-protein complex possesses two-fold rotational symmetry, which has been incorporated in the crystalline lattice.     

A comparison of electrophysiologically determined spectral responses in 35 species of Lepidoptera

Spectral responses from the compound eyes of 35 lepidopteran species representing 14 families were investigated electrophysiologically using ERG recordings. The light-stimuli used overed the range of 383–700 nm wavelengths. All species show three or four maxima in their spectral sensitivity curves. Two of these peaks were usually associated with ultraviolet and blue light (383 and 460 nm, respectively). The other maxima occurred in the 500–620 nm region. In Nymphalidae the highest peak was found in response to 560–580 nm stimuli. Of all wavelengths tested, these are the longest wavelengths to produce principal peak sensitivities.Pieridae and Lycaenidae have maxima in the UV region which represent significantly higher sensitivities than the secondary peaks to stimuli of longer wavelengths.Satyridae, Danaidae, Hesperiidae and diurnal moths except Epicopeia (Epicopeidae) generally have similar sensitivity curves with principal peaks between 500 and 520 nm.In Papilionid species except Graphium (max = 560 nm) high maxima occur in the UV and blue (460 nm) region.Noctural Sphingid moths possess the highest peak sensitivity at 540 nm. All other noctural moths tested have three or four maxima.     

Hybridization of synthetic oligodeoxyribonucleotides to phi chi 174 DNA: the effect of single base pair mismatch.

Oligodeoxyribonucleotides complementary to the DNA of the wild type (wt) bacteriophage phi chi 174 have been synthesized by the phosphotriester method. The oligomers, 11, 14, and 17 bases long, are complementary to the region of the DNA which accounts for the am-3 point mutation. When hybridized to am-3 DNA, the oligonucleotides form duplexes with a single base pair mismatch. The thermal stability of the duplexes formed between wt and am-3 DNAs has been measured. The am-3 DNA:oligomer duplexes dissociate at a temperature about 10 degrees C lower than the corresponding wt DNA:oligomer duplexes. This dramatic decrease in thermal stability due to a single mismatch makes it possible to eliminate the formation of the mismatched duplexes by the appropriate choice of hybridization temperature. These results are discussed with respect to the use of oligonucleotides as probes for the isolation of specific cloned DNA sequences.     

Amino acid sequence of galactosamine-containing glycopeptides in the hinge region of a human immunoglobulin D

Amino acid sequence and the location of seven galactosamine oligosaccharide moieties of the hinge region of the δ chain of human IgD NIG-65 have been determined. These oligosaccharide moieties are distributed in two distinct fashions: 1) three clusters each consisting of five amino acid residues with two consecutive attachment sites either Ala-X-Ala-Ser-Ser or Ala-X-Ala-Thr-Thr, where X can be any amino acid including proline, 2) one triplet sequence Val-Pro-Thr with one attachment site. We propose two rules with regard to the acceptor sequence for galactosamine oligosaccharides, the quintet sequence rule and triplet sequence rule.     

Carbon and nitrogen stable isotope ratios of deposit-feeding polychaetes in the Nanakita River Estuary, Japan

Two types of deposit-feeding polychaetes, Neanthes japonica and Notomastus sp., and their surrounding sediments were collected from the Nanakita River Estuary and a small brackish lagoon (Gamõ Lagoon) in northeastern Japan. The samples were examined using stable isotope analysis to assess the site specific feeding mode of the animals and their trophic status. N. japonica is a surface deposit-feeder and Notomastus sp. is a subsurface deposit-feeder. In the estuary, the sedimentary ⁵N tended to become isotopically heavier from the upper estuary (2.0 3.9) to the river mouth (4.3 6.2), while sedimentary organic ¹³C constant value (–26.8 –24.4, average –25.6) throughout the river estuary. The ¹³C values of N. japonica were similar to those of the surrounding sediment in the upper estuary, whereas in the lower estuary, N. japonica had a heavier ¹³C value than the surrounding sediment. The ¹³C and dg ¹⁵N values indicated that the carbon, but not the nitrogen, of N. japonica was derived from upland plants in the upper estuary. In the lower estuary, a significant fraction of carbon of N. japonica was derived from phytoplankton. Notomastus sp. exhibited heavier ¹³C values than the surrounding surface sediment throughout the estuary and had heavier ¹³C values than N. japonica in the same location. These results suggest selective utilization of sedimentary carbon by those animals following bacterial processing and subsequent fractionation. The difference in ¹⁵N between sedimentary organics and corresponding polychaetes was 5 ± 1 and rather higher than 3.4 ± 1.1 expected for normal trophic effects in other animals.     

Polo-like kinase 1 facilitates chromosome alignment during prometaphase through BubR1

Plk1, an evolutionarily conserved M phase kinase, associates with not only spindle poles but also kinetochores during prometaphase. However, the role of Plk1 at kinetochores has been poorly understood. Here we show that BubR1 mediates the action of Plk1 at kinetochores for proper chromosome alignment. Our results show that BubR1 colocalizes with Plk1 at kinetochores of unaligned chromosomes and physically interacts with Plk1 in prometaphase cells. Down-regulation of Plk1 by small interfering RNA abolished the mobility-shifted, hyperphosphorylated form of BubR1 in the prometaphase-arrested cells. In addition, BubR1 was phosphorylated by Plk1 in vitro at two Plk1 consensus sites in the kinase domain of BubR1. The add-back of either wild-type BubR1 or BubR1 2E, in which the two Plk1 phosphorylation sites were replaced by glutamic acids, but not that of BubR1 2A, an unphosphorylatable mutant, rescued the chromosome alignment defects in BubR1-deficient cells. Moreover, when both Plk1 and BubR1 were down-regulated, the add-back of BubR1 2E, but not that of wild-type BubR1, rescued the chromosome alignment defects. These results taken together suggest that Plk1 facilitates chromosome alignment during prometaphase through BubR1.     

Purification of phosphoproteins by immobilized metal affinity chromatography and its application to phosphoproteome analysis

Prefractionation procedures facilitate the identification of lower-abundance proteins in proteome analysis. Here we have optimized the conditions for immobilized metal affinity chromatography (IMAC) to enrich for phosphoproteins. The metal ions, Ga(III), Fe(III), Zn(II), and Al(III), were compared for their abilities to trap phosphoproteins; Ga(III) was the best. Detailed analyses of the pH and ionic strength for IMAC enabled us to determine the optimal conditions (pH 5.5 and 0.5 m NaCl). When whole cell lysates were fractionated in this way, about one-tenth of the total protein was recovered in the eluate, and the recovery of phosphorylated extracellular signal-regulated kinase (ERK) was more than 90%. Phosphorylated forms of ribosomal S6 kinase (RSK) and Akt were also enriched efficiently under the same conditions. Our Ga(III) IMAC and a commercially available purification kit for phosphoproteins performed similarly, with a slight difference in the spectrum of phosphoproteins. When phosphoproteins enriched from NIH3T3 cells in which ERK was either activated or suppressed were analyzed by two-dimensional fluorescence difference gel electrophoresis, phosphorylated ERK was detected as discrete spots unique to ERK-activated cells, which overlapped with surrounding spots in the absence of prefractionation. We applied the same technique to search for Akt substrates and identified Abelson interactor 1 as a novel potential target. These results demonstrate the efficacy of phosphoprotein enrichment by IMAC and suggest that this procedure will be of general use in phosphoproteome research.     

Expression of estrogen induced gene 121-like (EIG121L) during early Xenopus development

Estrogen induced gene 121 (EIG121) and EIG121-like (EIG121L) are evolutionarily conserved genes. But, their function is still unknown. Here, we report the expression pattern of Xenopus EIG121-like (xEIG121L) during early development. Its expression was first detected at stage 9 after mid-blastula transition, attained its maximal level at the gastrula stage, and remained constant until the tadpole stage. Whole-mount in situ hybridization revealed that xEIG121L was expressed strongly in the ventral ectoderm at the gastrula stage, and in the anterior ectoderm surrounding the neural plate at the neurula stage. xEIG121L expression was especially high in the presumptive hatching gland and cement gland regions in the neurula. At the tailbud stage, xEIG121L expression was limited to the hatching gland; an inverted Y type staining, characteristic of the hatching gland, was observed. However, at the tadpole stage, xEIG121L was expressed broadly in the head, heart and fin.