Cytochalasin D inhibits completion of cytokinesis and affects theca formation in dinoflagellates

Summary In the presence of cytochalasin D, dinoflagellates undergo mitosis and the cells begin to divide, but the completion of cell division is inhibited. InPausenella (dinospore formation),Gymnodinium andProrocentrum, Siamese twins arise which remain connected at the epicones whereas the hypocones, containing the nuclei, are separated. InScripsiella where the nucleus is centrally located, irregular binucleate cell bodies result. Cyst divisions which give rise to secondary or tertiary cysts inPaulsenella are not affected. In the athecatesPaulsenella andGymnodinium the morphogenesis of the separated cell portions is not or nearly not, respectively, disturbed by cytochalasin D. In the thecatesScripsiella andProrocentrum morphogenesis is heavily affected. InProrocentrum, wrinkled theca material is deposited instead of complete valvae. Doubling of the flagellar apparatus is not inhibited. It is concluded that the first phase of cytokinesis does not depend on actin. The daughter cells begin to separate by a mechanism which seems to be associated with the mitotic apparatus. Actin, however, is involved in the further constriction of the cleavage furrow in the second phase of cytokinesis and in the morphogenesis of the theca.     

The regulation of urease activity in Aspergillus nidulans

Aspergillus nidulans can utilize urea as a sole source of nitrogen but not as a carbon source. Urea is degraded by a urease. Mutation at any one of three genes, ureB, ureC, and ureD, may result in deficient urease activity. The ureB gene is closely linked to ureA, the structural gene for the urea transport protein. The heat lability of a ureB ^– revertant strain, intragenic complementation tests, and the linkage of ureB to ureA suggest that ureB is the urease structural gene. The ureD gene is probably involved in the synthesis or incorporation of a nickel cofactor essential for urease activity. The function of the ureC gene is not known. Urease is not induced but is subject to nitrogen regulation. The urease activities of ammonium-derepressed mutants show that the effector of nitrogen regulation is more likely to be glutamine than ammonium. When glutamine is present in the medium, urease appears to be inactivated by some means which does not involve a newly synthesized protease or a direct interaction between glutamine and urease.     

Gene expression,X-inactivation,and methylation during spermatogenesis: The case of Zfa,Zfx, and Zfy in mice

While it has become clear that X-inactivation in the female soma is complete in mouse (in contrast to being “patchy” in man), the degree of X-inactivation in the testes has not been ascertained. We have compared autosomal and X-linked zinc finger homolog expression and X-linked and Y-linked zinc finger homolog methylation in an attempt to elucidate this question. Using RTPCR, we have extended earlier studies of Zfx and Zfa expression in developing testes and find that Zfa expression starts at the time of X-inactivation while Zfx expression is continuous. Cell separation studies did not preclude continued expression of Zfx in adult germ cells. The methylation status of four CCGG residues in the Zfx promoter was studied using PCR bridging this region before and after DNA digestion with the isoschizomers Msp I and Hpa II, the latter being methylation sensitive. Hpa II resistant Zfx promoter DNA was found in all female tissues, but not in male tissues, including the testes. Previous studies have shown that Zfy is expressed at meiosis (like Zfa and unlike Zfx). Despite its expression, the Zfy gene is adjacent to, or contains, highly methylated CCGG sites since hybridization after Msp I digestion detected multiple small fragments that were not released after DNA digestion with Hpa II. Thus, Zfx is not methylated in sperm, while Zfy is, in contrast to their apparent patterns of expression. © 1993 Wiley-Liss, Inc.     

<Emphasis Type="Italic">O</Emphasis>-GlcNAcylation of the <Emphasis Type="Italic">Plum pox virus</Emphasis> capsid protein catalyzed by SECRET AGENT: characterization of <Emphasis Type="Italic">O</Emphasis>-GlcNAc sites by electron transfer dissociation mass spectrometry

The capsid protein of Plum pox virus (PPV-CP) is modified with O-linked β-N-acetylglucosamine (O-GlcNAc). In Arabidopsis thaliana this modification is made by an O-GlcNAc transferase named SECRET AGENT (SEC). Modification of PPV-CP by SEC is hypothesized to have a direct role in the infection process, because virus titer and rate of spread are reduced in SEC mutants. Previous studies used deletion mapping and site-directed mutagenesis to identify four O-GlcNAc sites on the capsid protein that are modified by Escherichia coli-expressed SEC. The infection process was not affected when two of these sites were mutated suggesting that O-GlcNAcylation of these sites does not have a significant role in the infection process or that a subset of the modifications is sufficient. Since it is possible that the mutational mapping approach missed or incorrectly identified O-GlcNAc sites, the modifications produced by E. coli-expressed SEC were characterized using mass spectrometry. O-GlcNAcylated peptides were enzymatically tagged with galactose, the products were enriched on immobilized Ricinus communis agglutinin I and sequenced by electron transfer dissociation (ETD) mass spectrometry. Five O-GlcNAc sites on PPV-CP were identified. Two of these sites were not identified in by the previous mutational mapping. In addition, one site previously predicted by mutation mapping was not detected, but modification of this site was not supported when the mutation mapping was repeated. This study suggests that mapping modification sites by ETD mass spectrometry is more comprehensive and accurate than mutational mapping.     

Phylogenetics of Sigmodontinae (Rodentia, Muroidea, Cricetidae), with special reference to the akodont group, and with additional comments on historical biogeography

This is the first cladistic analysis of sigmodontine rodents (Cricetidae, Sigmodontinae) based on nuclear and mitochondrial DNA sequences. Two most parsimonious cladograms (7410 steps in length; CI=0.199; RI=0.523) were discovered. Sigmodontinae appears well supported. Sigmodon is sister to the remaining living sigmodontines. It is shown that Euneomys is not a phyllotine and that the Reithrodon group is not monophyletic. Results corroborate that the abrothricines form a natural group that is not part of the akodont radiation. The akodontine tribe is well supported, and is composed of five main clades, whose limits and relationships are thoroughly discussed. For instance, the scapteromyines do not form a natural group and they fall within the akodontine clade. Additionally, I present some taxonomic judgments and comments on the historical biogeography of sigmodontines in the light of the newly discovered relationships. For example, five akodontine divisions are suggested: the Akodon, the Bibimys, the Blarinomys, the Oxymycterus, and the Scapteromys Divisions. It is shown that traditional hypotheses of sigmodontine historical biogeography are falsified by the recovered topology.     

Antigenic determinants distinctive of Methanospirillum hungatei and Methanogenium cariaci identified by monoclonal antibodies

Monoclonal antibodies were prepared against two species of Methanomicrobiaceae. Antibody 1A is specific for Methanospirillum hungatei strain JF1 and the determinant it recognizes is expressed on the surface of JF1 cells, where it is exposed and accessible to antibody. The determinant is found in a polypeptide (MW<12,000) in the sheath that covers the bacterial cell; it is not present in Methanospirillum hungatei strain GP1; and it is not expressed on the surface of whole cells of the other 24 methanogenic bacteria tested. It is therefore a marker of strain JF1, consequently, antibody 1A is potentially useful for tracking JF1 and fragments thereof in a variety of samples. Antibody 7A is specific for Methanogenium cariaci JR1c. It did not react with any other methanogen tested, not even with Mg. marisnigri or Ms. hungatei JF1, although these cross-react with Mg. cariaci if tested with polyclonal antisera. Therefore antibody 7A recognizes specifically a marker of Mg. cariaci JR1c.Abbreviations SIA slide immunoenzymatic assay - SDS-PAGE sodium dodecylsulfate polyacrylamide gel electrophoresis     

INTERACTIONS OF PHYTOPLANKTERS CULTURED FROM A POLLUTED SALINE LAKE,ONONDAGA LAKE,NEW YORK1,2,3

Two-membered cultures of 3 phytoplankters isolated from Onondaga Lake, grown in defined media under laboratory conditions, were analyzed by electronic particle counts and statistical methods to determine antagonistic, neutral, or stimulatory relationships. Over the range studied (exponential phase of growth, nutrients not limiting), growth of Staurastrum paradoxum is markedly reduced in the presence of Chlamydomonas sp. Growth of Chlamydomonas is not affected by presence of Staurastrum. The initial concentration of Staurastrum has an effect on its subsequent growth. The initial concentration of Chlamydomonas does not have an effect on its subsequent growth nor that of Staurastrum. These effects are not mediated by a filterable factor (unless highly labile) or by competition for a nutrient. They resemble, and perhaps describe, fluctuations of these species in the lake.     

The protein-encoding gene T-urf13 is not edited in maize mitochondria

RNA editing of T-urf13, a gene specific to the mitochondria of cytoplasmic male-sterile, type-T (cms-T) maize, and an adjacent, cotranscribed geneorf221, have been studied by cDNA sequencing. No editing was detected in 22 cDNA clones. This is the only report of a polypeptide-encoding gene in higher-plant mitochondria that is not edited. T-urf13 may not be edited because it is derived largely from the coding and flanking regions, which are rarely edited, of a ribosomal RNA gene.orf221 is edited; however, the similarity between the predicted amino acid sequences oforf221 incms-T and normal cytoplasms is not increased.     

Elf5 regulation in the trophectoderm

Mouse Elf5 is expressed exclusively in the trophectoderm from the late blastocyst stage to postgastrulation. We demonstrate here that the proximal promoter is used for trophectoderm expression but is not sufficient on its own. In transgenic assays, deletion of a differentially methylated region (DMR) within the promoter has no effect on the activation and maintenance of trophectoderm expression and does not result in ectopic activity. Two redundant enhancers drive Elf5 expression to the extraembryonic ectoderm and ectoplacental cone. The enhancers, located in the 5′ half of intron 1 and 3′ half of intron 2, require the presence of 1.8 kbp, although not the DMR, of the endogenous proximal promoter for optimal activity. These trophectoderm enhancers are mouse specific. A cattle Elf5 BAC reporter transgene is not expressed in mouse trophectoderm although it is expressed in skin, a known foetal domain of mouse Elf5 expression. The established importance of Elf5 for mouse trophectoderm at pre- and perigastrulation stages is not a conserved mammalian feature as Elf5 expression localises to embryonic as opposed to trophectodermal ectoderm in cattle.     

ATR and MKP1 play distinct roles in response to UV‐B stress in Arabidopsis

Ultraviolet‐B (UV‐B) stress activates MAP kinases (MAPKs) MPK3 and MPK6 in Arabidopsis. MAPK activity must be tightly controlled in order to ensure an appropriate cellular outcome. MAPK phosphatases (MKPs) effectively control MAPKs by dephosphorylation of phosphothreonine and phosphotyrosine in their activation loops. Arabidopsis MKP1 is an important regulator of MPK3 and MPK6, and mkp1 knockout mutants are hypersensitive to UV‐B stress, which is associated with reduced inactivation of MPK3 and MPK6. Here, we demonstrate that MPK3 and MPK6 are hyperactivated in response to UV‐B in plants that are deficient in photorepair, suggesting that UV‐damaged DNA is a trigger of MAPK signaling. This is not due to a block in replication, as, in contrast to atr, the mkp1 mutant is not hypersensitive to the replication‐inhibiting drug hydroxyurea, hydroxyurea does not activate MPK3 and MPK6, and atr is not impaired in MPK3 and MPK6 activation in response to UV‐B. We further show that mkp1 leaves and roots are UV‐B hypersensitive, whereas atr is mainly affected at the root level. Tolerance to UV‐B stress has been previously associated with stem cell removal and CYCB1;1 accumulation. Although UV‐B‐induced stem cell death and CYCB1;1 expression are not altered in mkp1 roots, CYCB1;1 expression is reduced in mkp1 leaves. We conclude that the MKP1 and ATR pathways operate in parallel, with primary roles for ATR in roots and MKP1 in leaves.     

Evidence for the Activation of NADH-cytochrome c Reductase during Germination of Lettuce

A very rapid increase in particulate cytochrome c reductase activity during the very early stages of germination of lettuce is demonstrated. The increase in activity does not parallel water uptake or the increase in cytochrome oxidase activity. The increase is reversible on drying of imbibed seeds and is not inhibited by cyanide. It is concluded that the increase in activity is due to some kind of activation process and not to de novo synthesis of protein. Possible mechanisms of activation were investigated. It was not possible to simulate the activation process in vitro, in the isolated particulate fraction.     

Revision of <Emphasis Type="Italic">Actinastrea</Emphasis>, the most common Cretaceous coral genus

The very common and species-rich Scleractinian genus Actinastrea (family Actinastraeidae, suborder Archeocaeniina) is revised on the basis of the type material of its type species and additional material from the type locality. A lectotype is designated for the type species. It was discovered that Jurassic to Early Cretaceous corals currently assigned to Actinastrea do not fit into the concept of this genus. These species belong to the genus Stelidioseris, which is also revised on the basis of the type of the type species, including designating a lectotype. These two genera are distinguished by various characteristics: septal external parts are swollen in Actinastrea but not in Stelidioseris, the costae are confluent in Stelidioseris but not in Actinastrea, the coenosteum is granulated in Actinastrea but narrow than in Actinastrea and only with costae in Stelidioseris. Actinastrea is restricted to the Late Cretaceous (Late Turonian—Maastrichtian), whereas Stelidioseris originates in the Jurassic and reaches into the Late Cretaceous, but is less common from the Turonian on.     

The Vibrio parahaemolyticus effector VopC mediates Cdc42‐dependent invasion of cultured cells but is not required for pathogenicity in an animal model of infection

Vibrio parahaemolyticus is a Gram‐negative marine bacterium that causes acute gastroenteritis in humans. The virulence of V. parahaemolyticus is dependent upon a type III secretion system (T3SS2). One effector for T3SS2, VopC, is a homologue of the catalytic domain of cytotoxic necrotizing factor (CNF), and was recently reported to be a Rho family GTPase activator and to be linked to internalization of V. parahaemolyticus by non‐phagocytic cultured cells. Here, we provide direct evidence that VopC deamidates Rac1 and CDC42, but not RhoA, in vivo. Our results alsosuggest that VopC, through its activation of Rac1, contributes to formation of actin stress fibres in infected cells. Invasion of host cells, which occurs at a low frequency, does not seem linked to Rac1 activation, but instead appears to require CDC42. Finally, using an infant rabbit model of V. parahaemolyticus infection, we show that the virulence of V. parahaemolyticus is not dependent upon VopC‐mediated invasion. Genetic inactivation of VopC did not impair intestinal colonization nor reduce signs of disease, including fluid accumulation, diarrhoea and tissue destruction. Thus, although VopC can promote host cell invasion, such internalization is not a critical step of the disease process, consistent with the traditional view of V. parahaemolyticus as an extracellular pathogen.     

Investigation of the functional properties and regulation of three glutamine synthetase-like genes in Streptomyces coelicolor A3(2)

Streptomyces coelicolor A3(2) has three additional glnA-type genes besides the glutamine synthetase genes glnA (encoding GSI) and glnII (encoding GSII). The aim of this work was to characterize their functional properties and regulation. Sequence analyses revealed that GlnA2, GlnA3, and GlnA4 are dissimilar to S. coelicolor GSI and lack highly conserved amino acid residues involved in catalysis. In heterologous expression experiments, glnA2, glnA3, and glnA4, in contrast to glnA and glnII, were not capable of complementing the l-glutamine auxotrophy of an Escherichia coli glnA mutant. The lack of a conserved sequence motif reflecting adenylylation control of enzyme activity suggests that GlnA2, GlnA3, and GlnA4 are not regulated via adenylyltransferase-mediated modification. In DNA-binding assays, the OmpR-like regulator of nitrogen metabolism GlnRII, which interacts with the glnA and glnII promoters, did not bind to the upstream regions of glnA2, glnA3, and glnA4. These findings support the conclusion that glnA2, glnA3, and glnA4 are not directly involved in l-glutamine synthesis and nitrogen assimilation and are not subject to nitrogen control in S. coelicolor. The glnA3 gene product is similar to FluG, which is required for asexual sporulation in Aspergillus nidulans. However, inactivation of glnA3 does not block morphological differentiation in S. coelicolor.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .     

A revised synopsis of the genus Atractomorpha Saussure, 1862 (Orthoptera:Pyrgomorphidae), with an account of the African aberrans - group

Recent work has shown that the preliminary revision of Atractomorpha by Banerjee & Kevan (1960), while basically not altogether unsatisfactory, is in need of considerable modification. The most important changes made include the recognition, as good species, of a number of taxa previously relegated to synonymy or to subspecific status. Two previously undescribed species have been discovered in the African a&errans-group, and two new subspecies are erected. The conclusions are largely based upon a study of the phallic structures, which had not previously been deemed satisfactory in providing diagnostic characters. The variability of these structures in the various species is illustrated, including that due to immaturity in two of them. The concealed female copulatory structures are also figured. A new key to species and subspecies (as well as an abridged key) is presented. New species are:A. occidentalis and A. orientalis. New subspecies are:A. sinensis montana and A. psittacina affinis. Species resurrected from synonymy are:A. rufopunctata Bolivar, A. himalayica Bolivar, A. angusta Karsch, and A. similis Bolivar, of which A. australiana Bolivar proves to be a synonym, and not a subspecies of A. crenaticeps (Blanchard). The other species re-elevated from subspecific rank is A. rhodoptera Karsch. Further changes in synonymy are as follows:A. lanceolata Bolivar is a synonym of A. burri Bolivar and not of A. rhodoptera; A. dentifrons Bolivar is a synonym of A. similis and not of A. crenaticeps; A. brevicornis (Thunberg) is removed from partial synonymy with A. lata (Motschoulsky). A. blanchardi Bolivar is not synonymous with A. crenulata (Fabricius) but is conspecific with A. acutipennis (Guérin-Méneville) and is here tentatively recognized as a widely distributed south-west Asiatic subspecies (in place of A. brevis Uvarov, which, as now understood, is restricted to south-west Arabia). A. crenulata apparently has a more restricted distribution than previously thought, being largely replaced or complemented by A, angusta, except in India and Ceylon. Neotypes are designated for A. acutipennis blanchardi and for Truxalis oceanicus Montrouzier (=A. crenaticeps). Information regarding the type specimens of all nominal species of Atractomorpha except A. nipponica Steinmann is given, and lectotype designations are made wherever necessary. Photographs of almost all primary types, except that of A. nipponica, that have not previously been so illustrated are present. This last species may be a synonym of A. rufopunctata.     

The phylogenetic position of the bumble bee inquiline Bombus inexspectatus and implications for the evolution of social parasitism

The high alpine European bumble bee Bombus (Thoracobombus) inexspectatus is one of three taxa in which obligate social parasitism has evolved in bumble bees. Until now, the phylogenetic placement of this species has not been analyzed quantitatively because it is rare in nature. Here a specimen of B. inexspectatus is sequenced for five genes to assess its phylogenetic position relative to other bumble bee species. Phylogenetic estimation of B. inexspectatus places this species within the subgenus Thoracobombus, as expected based on morphology. This provides strong support for the acquisition of social parasitism in B. inexspectatus separate from that found in the bumble bees of the subgenus Psithyrus and B. (Alpinobombus) hyperboreus. Furthermore, B. inexspectatus is not the sister taxon of its host but is a near relative, suggesting a loose but not strict adherence to Emery’s rule. B. inexspectatus is a sister taxon to a clade including B. veteranus, a frequent nest usurper that has been suggested to be a facultative social parasite. The phylogenetic placement of all bumble bee social parasites relative to their hosts is discussed.     

REDOX SENSITIVITY AND LIGHT MODULATION OF ENZYME ACTIVITY IN THE RHODOPHYTES GRACILARIA TIKVAHIAE AND CHONDRUS CRISPUS1

One of the cysteine residues believed to be necessary for reductive light activation is lacking in the only red algal NADP-linked glyceraldehyde-3-P dehydrogenases for which sequences are available, namely Gracilaria verrucosa (Hudson) Papenfuss and Chondrus crispus Stackhouse. Consistent with the mechanism of light modulation proposed for this enzyme, which involves reduction of domain movement-restricting disulfide bonds, it is not reductively activated in Chondrus crispus extracts, and it is not light-activated in whole cells or dithiothreitol (DTT) activated in extracts of the North American species Gracilaria tikvahiae McLachlan. Fructosebisphosphatase and glucose-6-P dehydrogenase, two enzymes for which sequence information from these algae is not yet available, are both activated in crude extracts by DTT treatment, but only fructosebisphosphatase is light-activated in intact Gracilaria.