High Affinity of the 5'-Upstream Region of a Winged Bean Chymotrypsin Inhibitor Gene for Nuclear Matrix

The 5'-upstream region of a winged bean chymotrypsin inhibitorgene (WCI-3b) was found to have a high affinity for nuclearmatrix. The region, named WCI-3b MAR (matrix attachment region),is highly A+T-rich and contains multiple sites interacting withnuclear matrix. A MAR was also found in the corresponding regionof the WCI-x gene, another active gene of the WCI family. SeveralMAR-binding proteins were detected in the wheat nuclear matrix. ⁴Present address: Friedrich Miescher Institute, P.O. Box 2543,CH-4002 Basel, Switzerland. ⁵Present address: Research Institute for Biological Sciences(RIBS), Kayo-cho, Jyobo-gun, Okayama, 716–1241 Japan.     

The Deduced Amino Acid Sequence of Phytochrome from Adiantum Includes Consensus Motifs Present in Phytochrome B from Seed Plants

A cDNA for the phytochrome of the fern Adiantum capillus-venerisL. was cloned and sequenced. The deduced phytochrome is 50{smalltilde}55% identical to phytochromes of seed plants, and 68%identical to Selaginella phytochrome. Regions resemble thosein previously characterized phytochromes from ferns, lower plantsand seed plants. ³Present address: Yamanouchi Pharmaceutical Co., Ltd., 21 Miyukigaoka,Tsukuba-shi, Ibaraki, 305 Japan ⁴Present address: Plant Growth Regulation Laboratory, The Instituteof Physical and Chemical Research (RIKEN), Hirosawa 2-1, Wako-shi,Saitama, 351-01 Japan ⁵Present address: Advanced Research Laboratory, Hitachi, Ltd.,Hatoyama, Saitama, 350-03 Japan     

Germ-Tube Formation is Independent of DNA Synthesis during Spore Germination in the Fission Yeast, Schizosaccharomyces pombe

Germ-tube emergence took place at almost the same time as DNAsynthesis during spore germination in the fission yeast, Schizosaccharomycespombe. Neither the emergence nor the elongation of the germ-tubeswas inhibited by hydroxyurea. Spores harboring a temperature-sensitivecdc 10 mutation produced germ-tubes even at a restrictive temperature,which indicates that germ-tube formation is not dependent onDNA synthesis. ¹ Present address: Department of Microbial Genetics, ResearchInstitute for Microbial Diseases, Osaka University, Yamada-Oka,Suita, Osaka 565, Japan. (Received June 3, 1982; Accepted July 5, 1982)     

Localization and Binding Characteristics of a High-Affinity Binding Site for N--Acetylchitooligosaccharide Elicitor in the Plasma Membrane from Suspension-Cultured Rice Cells Suggest a Role as a Receptor for the Elicitor Signal at the Cell Surface

A high-affinity binding site for N-acetylchitooligosac-chlarideelicitor was found to localize in the plasma membrane from suspension-culturedrice cells. Binding kinetics as well as the specificity of thisbinding site corresponded well with the behavior of the ricecells to the editor. These characteristics suggest that thebinding site represents a functional receptor for N-acetylchitooligosaccharideelicitor in rice. ²Present address: Okinawa Prefectural Livestock ExperimentalStation, 2009-5 Shoshi, Nakijin-son, Okinawa, 905-04 Japan. ³Present address: School of Hygiene and Public Health, The JohnsHopkins University, 615 North Wolfe Street, Baltimore, Maryland,21205 U.S.A. ⁴Present address: University of Tenessee, Microbiology, knoxville,Tennessee, 37996 U.S.A.     

Periplasmic location of dissimilatory nitrate and nitrite reductases in a denitrifying phototrophic bacterium, Rhodopseudomonas sphaeroides forma sp. denitrificans

The localization of dissimilatory nitrate and nitrite reductasesof a denitrifying phototrophic bacterium, Rhodopseudomonas sphaeroidesforma sp. denitrificans, was investigated. Nitrate and nitritereductases were located in the periplasmic space of the bacteriumgrown anaerobically in the presence of nitrate either in lightor in darkness. Chromatophores showed nitrate and nitrite reductaseactivities when dithionite-reduced benzyl viologen was an electrondonor; this suggests that the enzymes were trapped inside thevesicles. ¹Present address: Japanese Red Cross Central Blood Center, Hiroo4-1-31, Shibuyaku, Tokyo 150, Japan. ²Present address: Plant Growth Laboratory, University of California,Davis, California 95616, U.S.A. (Received November 7, 1979; )     

Molecular Cloning and Characterization of S-Adenosyl-L-Methionine:Scoulerine-9-O-Methyltransferase from Cultured Cells of Coptis japonica

S-Adenosyl-L-methionine : scoulerine-9-O-methyltransferase (SMT)catalyzes the transfer of the S-methyl group of S-adenosyl-L-methionineto the 9-hydroxyl group of scoulerine during the biosynthesisof berberine. We have isolated functionally active cDNA clones(pCJSMTs) from a cDNA library prepared from cultured cells ofCoptis japonica. The longest cDNA insert (pCJSMT1) had an openreading frame that encoded 351 amino acids, but the calculatedmolecular mass (38,364 Da) of the deduced product was slightlylower than the experimentally determined molecular mass of purifiedSMT. Rapid amplification of the 5' end of the cDNA indicatedthat the full-length cDNA of SMT consisted of 1,458 nucleotidesthat encoded 381 amino acids. When the full-length cDNA wasexpressed in E. coli, the molecular mass of the expressed SMTwas greater than that of native SMT in Coptis cells. This resultsuggests that SMT might be produced in a pre-mature form andprocessed post-translationally. SMT was also found to exhibitsequence homology to other O-methyltransferases from plantsand N-terminal region of the SMT polypeptide appeared to benecessary for enzymatic activity. ¹Present address: High Quality Life Research Laboratories, SumitomoMetal Industries, Ltd., 3-5 Hikaridai, Seika, Sourakugun, Kyoto,619-02 Japan ²Present address: Suntory Research Center, 1-1-1 Wakayamadai,Shimamoto, Mishima-gun, Osaka, 618 Japan ³Present address: Department of Cell Biology, The Scripps ResearchInstitute, La Jolla, CA 92037 U.S.A.     

Analysis of Lipids in Prochloron sp.: Occurrence of Monoglucosyl Diacylglycerol

Prochloron contained monogalactosyl diacylglycerol, digalactosyldiacylglycerol, sulfoquinovosyl diacylglycerol, phosphatidylglyceroland, as a minor component, monoglucosyl diacylglycerol, butno phosphatidylcholine. With respect to the lipid and fattyacid compositions, this alga is similar to the blue-green algaerather than the chloroplasts of eukaryotic plants. ¹ Present address: Department of Biology, Faculty of Science,University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113, Japan. (Received August 24, 1982; Accepted November 18, 1982)     

Photoconversion of protochlorophyll to chlorophyll a in a mutant of Chlorella regularis

Dark-grown cells of a mutant strain of Chlorella regularis containedchlorophyll a and protochlorophyll, phytyl ester of protochlorophyllide.Under illumination, protochlorophyll was quantitatively anddirectly converted into chlorophyll a. The photoconversion wasdependent on light intensity and temperature and proceeded ina cell-free preparation. The pathway of chlorophyll formation found in the mutant cellsis entirely different from that from protochlorophyllide byway of chlorophyllide a, which is generally observed in greenplants. ¹Present address: Division of Biology, Medical College of Miyazaki,Miyazaki 889-16, Japan. ²Present address: Division of Environmental Biology, The NationalInstitute for Environmental Studies, Ibaragi 300-21, Japan. (Received October 24, 1975; )     

Induction of Cytosolic Acetyl-Coenzyme A Carboxylase in Pea Leaves by Ultraviolet-B Irradiation

Levels of subunits of two acetyl-coenzyme A carboxylases werehigh in small leaves of Pisum sativum, decreased with growth,and remained constant in fully expanded leaves. Irradiationof fully expanded leaves induced the cytosolic isozyme only.This result suggests a key role for the cytosolic enzyme inprotection against UV-B. ¹Present address: Laboratory of Molecular Genetics, BiotechnologyInstitute, Akita Prefectural College of Agriculture, 2-2 Minami,Ohgata, Akita, 010-04 Japan ²Present address: Laboratory of Plant Molecular Biology, Schoolof Agricultural Sciences Nagoya University, Nagoya, 464-01 Japan     

Studies on Microbody Development in Wounded Sweet Potato Root Tissue: Proposal for the Post-Translational Transport of Catalase into Preexisting Microbodies

Pure microbody fractions could be prepared in considerable yieldsfrom sweet potato root tissue slices incubated for 16 hr and3 days. The ratio of catalase activity to phospholipid contentin the fraction from slices incubated for 3 days was about 3times that from slices incubated for 16 hr. Total catalase activityin the former slices was about twice that in the latter. Thissuggests that catalase synthesized during incubation of theslices is transported into microbodies preexisting in intacttissue. ¹ Present address: Laboratory of Food Technology, Faculty ofApplied Biological Science, Hiroshima University, Fukuyama,Hiroshima 720, Japan. ² Present address: Terumo Co., Ltd., Omiya, Fujinomiya, Shizuoka418, Japan. (Received July 1, 1982; Accepted September 24, 1982)     

Nitrite-reducing activity of modified cytochrome c-553 from the red alga Porphyra yezoensis Ueda

Crystalline cytochrome c-553 was obtained from Porphyra yezoensisUeda. The cytochrome in areduced form was modified to show anitrite-reducing activity after appropriate treatment with heat,hydrogen peroxide, or photooxidation using methylene blue asthe electron acceptor, but the reducing activity was far lowerthan that of the nitrite reductase isolated from this alga.The modified cytochrome c-553 was autooxidizable and showedan absorption spectrum resembling that of cytochrome c-553 inthe oxidized form except for slight shifts of the absorptionmaximumin the -band region toward shorter wavelengths. ¹ Present address: Department of Biological Sciences, Universityof Tsukuba, Sakura-Mura, Ibaraki, 300-31 Japan. ² Present address: Department of Fisheries, College of Agricultureand Veterinary Medicine, Nihon University, Shimouma, Setagaya-ku,Tokyo, 154 Japan. (Received June 10, 1975; )     

Purification and Some Properties of Cytochrome b-560 from Nitrosomonas europaea

Cytochrome b-560 was purified to an electrophoretically homogeneousstate from Nitrosomonas europaea. It showed absorption peaksat 427, 530 and 560 nm in the reduced form. Its molecular weightwas estimated to be 44,000 by SDS-polyacrylamide gel electrophoresisand the same value was obtained on the basis of the contentsof haem and protein. The cytochrome was not autoxidizable anddid not react with CO. ¹Present address: Tokyo Research Center, TOSOH Corporation,Hayakawa, Ayase-shi, Kanagawa 252, Japan ²Present address: Faculty of Integrated Arts and Sciences, HiroshimaUniversity, Higashisenda-machi, Hiroshima 730, Japan (Received March 23, 1988; Accepted June 2, 1988)     

Phenotypic Characterization and Molecular Mapping of an acaulis2 Mutant of Arabidopsis thaliana with Flower Stalks of Much Reduced Length

An acaulis2-1 (acl2-1) mutant of Arabidopsis thaliana (L.) Heynh.was isolated and characterized. The mutant had inflorescencesof much reduced length. The defect was severer in lower internodesand was visible preferentially in type 2 metamers. The defectin the elongation of internodal cells of inflorescences wasattributable to a defect in the mechanism for elongation ofthese cells. The cortical microtubules (MTs) were analyzed inthe acl2 mutant, in a comparison with those of an acl1 mutantwhich was described earlier, but no changes were detected inamounts of MTs. In the type 2 and type 3 metamers, the defectin elongation in acl mutants was loosely correlated with a changein the orientation of MTs. Thus, the abnormal arrangement ofMTs appears to be one of the pleiotropic effects of acaulismutations. Genetic mapping was performed using molecular markers and theACL2 gene was located at 100 map units on chromosome 1. ¹Present address: Institute of Molecular and Cellular Biosciences,The University of Tokyo, Yayoi, Bunkyo-ku, Tokyo, 113 Japan ²Present address: Division of Biological Sciences, GraduateSchool of Science, Hokkaido University, Kita-ku, Sapporo, 060Japan     

Studies on anaerobic biphasic growth of a denitrifying bacterium, Pseudomonas stutzeri

Growth of Pseudomonas stutzeri(VAN NIEL strain) in the presenceof a limiting amount of nitrate under anaerobic conditions ischaracterized by 2 logarithmic phases separated distinctly byan intermediate phase where the growth rate is very low. Inthe first logarithmic phase nitrate is reduced stoichiometricallyto nitrite stage, and in the second phase nitrite is reducedto nitrogen gas. The nitrite reducing activity of cells in the second growthphase is 3–4 times higher than that of cells in the firstphase. The rise in nitrite reducing activity is correlated witha remarkable increase in the content of cytochromes a₂ and c-552. ¹Present address: Department of Biochemistry, Hiroshima UniversitySchool of Dentistry, Hiroshima, Japan. ²Present address: Institute of Molecular Biology, Faculty ofScience, Nagoya University, Nagoya, Japan. (Received June 16, 1969; )     

Tentoxin Inhibits Both Photophosphorylation in Thylakoids and the ATPase Activity of Isolated Coupling Factor F1 from the Cyanobacterium Anacystis nidulans

Tentoxin strongly inhibited the ATPase activity of isolatedcoupling factor 1 (AF₁) from the cyanobacterium Anacystis nidulans,with 50% inhibition occurring at 0.3 µM. When thylakoidsfrom A. nidulans were preincubated with 0.3 µM tentoxinfor 30 min, photophosphorylation was inhibited by 50%. Measurementsof fluorescence from 9-aminoacridine indicated that tentoxininhibited the utilization of the proton gradient by ATP formationin thylakoids. These results indicate that tentoxin is a strongenergy-transfer inhibitor of photophosphorylation in A. nidulans.Tentoxin decreased the level of ATP in intact cells both inthe light and in darkness, its effects being much stronger inthe dark. Tentoxin at 50 µM strongly inhibited the growthof the cells. ³Present address: Corporate Research and Development Laboratory,Tonen Co. 1-3-1 Nishi-tsurugaoka, Ohi-machi, Saitama, 354 Japan ⁴Present address: Technology and Engineering Laboratories, AjinomotoCo., Inc. Suzuki-cho 1, Kawasaki, 210 Japan     

Metabolism of p-Hydroxybenzoate via Hydroxyquinol by Trichosporon cutaneum WY2-2: Characterization of the Pathway using Superoxide Dismutase as a Stabilizer of Hydroxyquinol

Trichosporon cutaneum WY2-2 was shown to metabolize p-hydroxybenzoatevia protocatechuate and hydroxyquinol. Using superoxide dismutaseas a stabilizer of hydroxyquinol, the conversion of protocatechuateto hydroxyquinol and the ring fission process of hydroxyquinolwere confirmed. Hydroxyquinol was chemically identified as theproduct of protocatechuate hydroxylase reaction. Partially purifiedprotocatechuate hydroxylase was highly specific for protocatechuate;its K_m values for protocatechuate and NADH were 17.6 and 12.4µM, respectively. It catalyzed equimolar CO₂ formation,NADH oxidation and O₂ consumption from protocatechuate. Hydroxyquinoldioxygenase was highly specific for hydroxyquinol, with a K_mof 2.9 µM. ¹A preliminary account of this work was presented at the 81stMeeting of the Chubu-branch of Agricultural Chemical Societyof Japan, Gifu, October, 1980. ²Present address: Biological Institute, Faculty of Science,Nagoya University, Nagoya 464, Japan. ³Present address: Shin Nihon Chemical Co. Ltd... 19-10, Showa-cho,Anjoh, Aichi 446, Japan. (Received November 15, 1985; Accepted August 27, 1986)     

Demonstration of light-induced potential change in Chara cells lacking tonoplast

Chara cells without tonoplasts, prepared by replacing the cellsap with EGTA-containing media, showed essentially the samepattern of light-induced changes in membrane potential and membraneresistance as normal cells although the concentrations of ionsand ATP in the cytoplasm decreased considerably (1/3–1/10)after loss of the tonoplast. Removal of the tonoplast reducedthe rate of photosynthetic O₂ evolution to about 50% of thatof normal cells but did not affect the magnitude of light-inducedpotential change. Not a full but a certain level of electronflow seems necessary to activate the putative electrogenic H⁺-pump. ¹ Present address: Department of Botany, Faculty of Science,University of Tokyo, Japan. ² Present address: Niigata College of Pharmacy, Niigata 950-21,Japan. (Received September 4, 1978; )     

Separation and Partial Characterization of Membranes from Prochloron sp.

Two differently colored membrane preparations were separatedfrom the prochlorophyte, Prochloron sp., by mechanical disintegrationof the cells followed by sucrose density gradient centrifugation.An orange-colored preparation, containing zeaxanthin as themajor constituent pigment, seemed to comprise the cytoplasmicmembrane. The other green-colored membrane preparation, containingß-carotene and chlorophyll a and b as major pigmentconstituents, was identified as the thylakoid membrane. Thetwo types of membranes were compared as to their absorptionspectra and buoyant densities. ¹ This work is one of the results of the 8th International Expeditionon Prochloron organized by Dr. R. A. Lewin, University of Californiaat San Diego. ⁵ Present address: Solar Energy Research Group, The Algatron,The Institute of Physical and Chemical Research (RIKEN), Wako-shi,Saitama 351, Japan. ⁶ Present address: National Institute for Basic Biology, Okazaki444, Japan. (Received October 19, 1984; Accepted January 7, 1985)