Slipped-strand mispairing in a plastid gene: rpoC2 in grasses (Poaceae)

An exception to the generally conservative nature of plastid gene evolution is the gene coding for the beta" subunit of RNA polymerase, rpoC2. Previous work by others has shown that maize and rice have an insertion in the coding region of rpoC2, relative to spinach and tobacco. To assess the distribution of this extra coding sequence, we surveyed a broad phylogenetic sample comprising 55 species from 17 angiosperm families by using Southern hybridization. The extra coding sequence is restricted to the grasses (Poaceae). DNA sequence analysis of 11 species from all five subfamilies within the grass family demonstrates that the extra sequence in the coding region of rpoC2 is a repetitive array that exhibits more than a twofold increase in nucleotide substitution, as well as a large number of insertion/deletion events, relative to the adjacent flanking sequences. The structure of the array suggests that slipped-strand mispairing causes the repeated motifs and adds to the mechanisms through which the coding sequence of plastid genes are known to evolve. Phylogenetic analyses based on the sequence data from grass species support several relationships previously suggested by morphological work, but they are ambiguous about broad relationships within the family.      

Genetic variability and differentiation in the larch populations within the range of <Emphasis Type="Italic">Larix olgensis</Emphasis> A. Henry in Primorye

The level of within-and among-population variation of larch inhabiting the range of Larix olgensis A. Henry in Primorye was estimated on the basis of 440 RAPD loci identified by means of 12 random primers. In ten populations examined, the proportion of polymorphic loci was 35–60%, the average expected heterozygosity varied from 0.1340 to 0.2169, and the average gene flow estimate was 1.38. According to Fisher’s test for heterogeneity, the pairwise differences of the fragment frequencies between the populations were statistically significant. The subdivision index G _ST = 0.2663 indicated that the interpopulation variation component accounted for approximately 27% of the total variation. Coefficients of Nei’s genetic distance between the populations varied from 0.0137 to 0.0934. Most of the samples with high support clustered according to the geographic positions relative to one another within the range. These results suggest that the populations examined are characterized by high genetic variation, like the larch populations of Siberia and the Russian Far East studied earlier, but, in contrast to the latter, exhibit higher among-population variability.     

Bacteria of the <Emphasis Type="Italic">Burkholderia cepacia</Emphasis> complex are cyanogenic under biofilm and colonial growth conditions

Background  

The Burkholderia cepacia complex (Bcc) is a collection of nine genotypically distinct but phenotypically similar species. They show wide ecological diversity and include species that are used for promoting plant growth and bio-control as well species that are opportunistic pathogens of vulnerable patients. Over recent years the Bcc have emerged as problematic pathogens of the CF lung. Pseudomonas aeruginosa is another important CF pathogen. It is able to synthesise hydrogen cyanide (HCN), a potent inhibitor of cellular respiration. We have recently shown that HCN production by P. aeruginosa may have a role in CF pathogenesis. This paper describes an investigation of the ability of bacteria of the Bcc to make HCN.     

Pregna-D′-pentaranes,Progestins and Antiprogestins: II. Pathways and Realization Mechanisms of Separate Biological Functions of Steroid Hormones

The manner in which multifunctional steroid hormones realize their separate biological functions in mammal organisms is considered. This study is carried out on the basis of a systematic set of progesterone analogues, which we developed and described in part I of the review. This set has for the first time enabled the use of compounds of this type for studying the pathways and realization mechanisms of separate biological functions of steroid hormones. The interaction of pregna-D′-pentaranes with the classical progesterone receptor, their independent influence on the myometrium and ovogenesis, and some nonclassical effects are described. A scheme of realization is suggested for the biological functions already known, newly discovered, and presumed by us that, during pregnancy, are fulfilled in mammal organisms by progesterone and its nearest metabolite, dihydroprogesterone.__________Translated from Bioorganicheskaya Khimiya, Vol. 31, No. 3, 2005, pp. 227–238.Original Russian Text Copyright © 2005 by Kamernitzky, Levina.The Cycle of Works of the Authors Described in the Review Has Won the Shemyakin Prize in 2004For part I, see [1].     

Steroid compounds from far Eastern starfishes Henricia aspera and H. tumida

Six new natural compounds were isolated from two Far Eastern starfish species, Henricia aspera and H. tumida, collected in the Sea of Okhotsk. Two new glycosylated steroid polyols were obtained from H. aspera: asperoside A and asperoside B, which were shown to be (20R,24R,25S)-3-O-(2,3-di-O-methyl-beta-D-xylopyranosyl)-24-methyl-5alpha-cholest-4-ene-3beta,6beta,8,15a,16beta,26-hexaol and (20R,24R,25S,22E)-3-O-(2,4-di-O-methyl-beta-D-xylopyranosyl)-24-methyl-5alpha-cholest-22-ene-3beta,4beta,6beta,8,15alpha,26-hexaol, respectively. Two other glycosylated polyols, tumidoside A, with the structure elucidated as (20R,22E)-3-O-(2,4-di-O-methyl-beta-D-xylopyranosyl)-26,27-di-nor-24-methyl-5alpha-cholest-22-ene-3beta,4beta,6beta,8,15alpha,25-hexaol, and tumidoside B, whose structure was elucidated as (20R,24S)-3-O-(2,3-di-O-methyl-beta-D-xylopyranosyl)-5alpha-cholestan-3beta,4beta,6beta,8,15alpha,24-hexaol, were isolated from the two starfish species. (20R,24S)-Salpha-Cholestan-3beta,6beta,15alpha,24-tetraol and (20R,24S)-5alpha-cholestan-3beta,6beta,8,15alpha,24-pentaol were identified only in H. tumida. The known monoglycosides henricioside H1 and laeviuscolosides H and G were also identified in both species.     

Pregna-D'-pentaranes, progestins, and antiprogestins: II. Pathways and realization mechanisms of separate biological functions of steroid hormones

The manner in which multifunctional steroid hormones realize their separate biological functions in mammal organisms is considered. This study is carried out on the basis of a systematic set of progesterone analogues, which we developed and described in part I of the review. This set has for the first time enabled the use of compounds of this type for studying the pathways and realization mechanisms of separate biological functions of steroid hormones. The interaction of pregna-D'-pentaranes with the classical progesterone receptor, their independent influence on the myometrium and ovogenesis, and some nonclassical effects are described. A scheme of realization is suggested for the biological functions already known, newly discovered, and presumed by us that, during pregnancy, are fulfilled in mammal organisms by progesterone and its nearest metabolite, dihydroprogesterone. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 3; see also http://www.maik.ru.     

Cytoskeletal control of cell length regulation

It was shown that mouse embryo fibroblasts and human foreskin diploid fibroblasts of AGO 1523 line cultivated on specially prepared substrates with narrow (15 +/- 3 microns) linear adhesive strips were elongated and oriented along the strips, but the mean lengths of the fibroblasts of each type on the strips differed from those on the standard culture substrates. In contrast to the normal fibroblasts, the length of mouse embryonic fibroblasts with inactivated gene-suppresser Rb responsible for negative control of cell proliferation (MEF Rb-/-), ras-transformed mouse embryonic fibroblasts (MEF Rb-/-ras), or normal rat epitheliocytes of IAR2 line significantly exceeded those of the same cells on the standard culture substrates. The results of experiments with the drugs specifically affecting the cytoskeleton (colcemid and cytochalasin D) suggest that the constant mean length of normal fibroblasts is controlled by a dynamic equilibrium between two forces: centripetal tension of contractile actin-myosin microfilaments and centrifugal force generated by growing microtubules. This cytoskeletal mechanism is disturbed in MEF Rb-/- or MEF Rb-/-ras, probably, because of an impaired actin cytoskeleton and also in IAR2 epitheliocytes due to the different organization of the actin-myosin system in these cells, as compared to that in the fibroblasts.     

Morphological Changes in Rat Uterus Tissues under the Action of Progestins and Antiprogestins of the Pregna-D"-pentaran Series

Changes in the uterus morphology of mature female rats were studied on the model of pseudopregnancyafter treatment with the progestin 16,17-cyclohexanoprogesterone (PR) and the antirpogestins 5(H)-16,17-cyclohexano-4,5-dihydroprogesterone (APR1) and 5(H)-16,17-cyclohexano-4,5-dihydroprogesterone (APR2). The rats were preliminarily estrogenized with 17-estradiol at a dose of 1 g/(animal day) for 4 days and then treated with PR at a dose of 0.2 mg/(animal day) for 14 days. The first group was then left without any treatment, whereas APR1 and APR2 were injected at the dose of 0.2 mg/(animal day) for 4 days to the animals of the second and the third groups, respectively. Light and electron microscopy of the uterus preparations demonstrated that the PR action provoked a complete pseudopregnancy picture characterized by the endometrium functionalization and the myometrium hypertrophy. Subsequent treatment with APR1 and APR2 caused the hypertrophy to cease, which had a more pronounced effect in the case of APR2. At the same time, some indications of the endometrium functionalization remained observable after treatment with APR1 and APR2. The specific binding sites for ³H-labeled APR1 and APR2 were absent from the uterus cytosol for the rats gestagenized with PR.     

Genetic variability and interrelationship of Siberian and Far Eastern larches from RAPD-analysis data

Genetic diversity of larches from six geographically isolated regions, Tomsk, Irkutsk, Ulan-Ude (Siberia), and Blagoveshchensk, Khabarovsk, Yuzhno-Sakhalinsk (Far East) was examined by means of RAPD analysis. Tree DNA samples were compared using 457 RAPD loci (97% of which were polymorphic), identified with 17 primers of random sequences. In the samples examined, 32 to 49% of the genes were in heterozygous state, mean expected heterozygosity (Hexp) varied from 0.1373 to 0.1891, and the genetic distances (DN) for different sample pairs varied from 0.0361 to 0.1802. The main population parameters were determined for Larix sibirica Ledeb., L. gmelinni (Rupr.) Rupr., and L. kamtschatica (Rupr.) Carr. Analysis of the genetic relationships showed that L. kamtschatica was characterized by highest genetic differentiation from the other larches examined, while larches from Primorskii krai were genetically close to L. sibirica.     

Gadolinium Effects on Gigaseal Formation and the Adhesive Properties of a Fungal Amoeboid Cell,the <Emphasis Type="BoldItalic">Slime</Emphasis> Mutant of <Emphasis Type="Italic">Neurospora crassa</Emphasis>

Low gadolinium concentrations induce rapid gigaseal formation and cell adhesion to glass and plastic (polystyrene) substrates in the slime mutant of Neurospora crassa. Cellular adhesion is independent of an integrin-mediated mechanism, because pretreatment with the oligopeptide ARG-GLY-ASP-SER (RGDS) did not inhibit it, and there was no spatial correlation between integrin and adhesions. In contrast, concanavalin A and beta-galactosidase both inhibit adhesion, suggesting that adhesion is mediated by sugar moeities at the cell surface. The adhesion sites are motile in the plasma membrane, as shown by the movement of polystyrene microspheres on the cell surface. In addition to an integrin-based adhesive system, which has already been characterized in walled hyphal cells, hyphae have evolved at least two different plasma membrane-based adhesion mechanisms. The relatively non-specific sugar-mediated adhesion caused by gadolinium may be part of the mechanism of gigaseal formation in other cells. In the absence of sugar-mediated adhesion, gadolinium increases the magnitude of the gigaseal in giant unilamellar liposomes composed of phosphatidylcholine, phosphatidylethanolamine, and cholesterol, with or without the negatively charged phosphatidylserine. Thus, gigaseal formation involves at least two different mechanisms. Abbreviations: BTP, bis-tris-propane; MES, morpholinoethanesulfonic acid; RGDS and RGDE, oligopeptides ARG-GLY-ASP-SER and ARG-GLY-GLU-SER, respectively; BS, bath solution; BSA, bovine serum albumin; PBS, phosphate-buffered saline; GULs, giant unilamellar liposomes; PC, Palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine; PE, 1-Palmitoyl-2-linoleoyl-sn-glycero-3-phosphoethanolamine; PS, 1-Palmitoyl-2-linoleoyl-sn-glycero-3-[phospho-L-serine] (sodium salt); Ch, Cholesterol.