Genetic control of the oil content in Thymus vulgaris L: a case of polymorphism in a biosynthetic chain

Six distinet chemotypes of the mediterranean Thymus vulgaris L. are characterized by differences in the major monoterpenes produced. The nature of the monoterpenes is shown to be controlled by a series of loci. Each locus probably has a dominant and a recessive allele. Epistatic relationships between these loci are interpreted by assuming that terpenes are end products of branches from the same biosynthetic pathway. The most dominant types, which correspond to the monoterpenes produced at the beginning of the chain, show the most important potential genetic diversity. There is good evidence that regulatory genos are involved in this system.     

The presence of chlorophyll b and the estimation of phaeopigments in marine phytoplankton

A reverse-phase h.p.l.c. technique was used to estimate theconcentration of chlorophyll b in phytoplankton cultures, fecalpellets of Calanus pacificus, and suspended paniculate matterfrom the Central North Pacific, Oregon coastal waters, and DabobBay (a temperate fjord in Puget Sound, WA, USA). The purposewas to assess the distribution of this pigment in the euphoticzone and its effect on the fluorometnc estimation of phaeopigments.Analyses of natural waters confirm high chlorophyll b concentrations(median mass ratio of b:a > 0.3) at the depth of the chlorophylla maximum in tropical waters while values for temperate planktonare relatively low (median mass ratio of chl b:a = 0.05) andpatchy. Zooplankton fecal pellets showed a significant enrichmentin chlorophyll b, suggesting grazing as a mechanism to explainhigh concentrations of this pigment at the bottom of the euphoticzone. It is estimated that the presence of chlorophyll b couldcause an average overestimation of phaeopigment concentrationby the fluorometnc technique of 38% between 0 and 200 m in theCentral North Pacific. This effect is more pronounced at thelayer of chlorophyll b maximum (120–140 m). ¹Present address: Marine Biology Research Division, A-002, ScrippsInstitution of Oceanography, La Jolla, CA 92093, USA     

Relationships of the Col plasmids E2, E3, E4, E5, E6, and E7: Restriction mapping and colicin gene fusions

Thirteen ColE plasmids representing the E2-E7 types have been compared by restriction mapping. Over 80% of their restriction sites were found to be similarly positioned, indicating that these plasmids share a common structure. Three variants are ColE2-CA42 and ColE7-K317, both of which contain 1.8-kb DNA segments in place of a 2.5-kb segment common to the other plasmids, and ColE6-CT14, which has an additional 5.0-kb DNA segment compared to the other plasmids. The colicin (col), immunity (imm), and colicin release (hic) genes of these plasmids have been localized to regions corresponding to those known for ColE3-CA38 and ColE2-P9, with the imm and hic genes adjacent to the 3' end of the col gene. Active colicin is produced from hybrid col genes containing 5' and 3' ends from different E-type plasmids. The 3'-termini of the fused col genes specify the colicin type.     

Evolutionary study of multigenic families mapping close to the human MHC class I region

Summary During a search for novel coding sequences within the human MHC class I region (chromosome 6p21.3), we found an exon (named B30-2) coding for a 166-amino-acid peptide which is very similar to the C-terminal domain of several coding sequences: human 52-kD Sjögren's syndrome nuclear antigen A/Ro (SS-A/Ro) and ret finger protein (RFP), Xenopus nuclear factor 7 (XNF7), and bovine butyrophilin. The first three of these proteins share similarities over the whole length of the molecule whereas butyrophilin is similar in the C-terminal domain. The N-terminal domain of butyrophilin is similar to rat myelin/oligodendrocyte glycoprotein (MOG) and chicken B blood group system (B-G) protein. These domains are components of a new subfamily of the immunoglobulin superfamily (IgSF). Butyrophilin is thus a mosaic protein composed of the MOG/B-G Ig-like domain and the C-terminal domain of 52-kD SS-A/Ro, RFP, and XNF7 (1330-2-like domain). Moreover, in situ hybridization shows that RFP, butyrophilin, and MOG map to the human chromosome 6p2l.3-6p22 region and are thus close to the MHC class I genes. It is therefore possible that the butyrophilin gene is the product of an exon shuffling event which occurred between ancestors of the RFP and MOG genes. To our knowledge, this is the first example of the colocalization of a chimeric gene and its putative progenitors. Finally, regulatory protein T-lymphocyte 1 (Rpt-1) shares similarities with the N-terminal halves of RFP, 52-kD SS-A/Ro, and XNF7, but not with the B30-2-like domain. We show that the ancestral Rpt-l gene evolved by overprinting. Correspondence to: P. Pontarotti     

Expression of the Saccharomyces cerevisiae Kex2p endoprotease in inset cells. Evidence for a carboxy-terminal autoprocessing event.

The pheromone-processing Kex2p endoprotease of Saccharomyces cerevisiae has been difficult to characterize due to its low level of expression in yeast cells. To overcome this problem, we have overexpressed Kex2p using the baculovirus/insect cell expression system. Spodoptera frugiperda Sf9 insect cells infected with a recombinant baculovirus, containing the complete KEX2 gene which encodes the Kex2p protease (814 amino acids), accumulate an 120-kDa functional form of the enzyme. The inhibition profile of the insect-cell-derived endoprotease is similar to that of the yeast enzyme. The recombinant infected insect cells also secrete into the medium about half of the total Kex2p activity produced. Deleting the carboxyl-terminal tail and the transmembrane domain of Kex2p (Kex2 delta p, 666 amino acids) does not measurably interfere with the enzyme characteristics and results in the secretion of up to 90% of the total enzyme activity. The truncated form, Kex2 delta p, of the endoprotease accumulates in the cell supernatant to 6.7 x 10(5) U/l. The molecular mass of the secreted forms for both the wild-type Kex2p and Kex2 delta p is the same (70 kDa) and is 50-kDa lower than the intracellular form. This result implicates a processing event which gives rise to shorter extracellular forms of both the wild-type Kex2p and Kex2 delta p and which trims their carboxy termini upsteam of amino acid 666. This processing event requires the integrity of the Ser385 of the Kex2p active site.     

Importance of hydrogen-bonding interactions involving the side chain of Asp158 in the catalytic mechanism of papain

In a previous study, it was shown that replacing Asp158 in papain by Asn had little effect on activity and that the negatively charged carboxylate of Asp158 does not significantly stabilize the active site thiolate-imidazolium ion pair of papain (Ménard et al., 1990). In this paper, we report the kinetic characterization of three more mutants at this position: Asp158Gly, Asp158Ala, and Asp158Glu. From the pH-activity profiles of these and other mutants of papain, it has been possible to develop a model that enables us to dissect out the contribution of the various mutations toward (i) intrinsic activity, (ii) ion pair stability, and (iii) the electrostatic potential at the active site. Results obtained with mutants that place either Gly or Ala at position 158 indicate that the hydrogen bonds involving the side chain of Asp158 in wild-type papain are indirectly important for enzyme activity. When CBZ-Phe-Arg-MCA is used as a substrate, the (kcat/KM)obs values at pH 6.5 are 3650 and 494 M-1 s-1 for Asp158Gly and Asp158Ala, respectively, as compared to 119,000 M-1 s-1 for papain. Results with the Asp158Glu mutant suggest that the side chain of Glu moves closer to the active site and cannot form hydrogen bonds similar to those involving Asp158 in papain. From the four mutations introduced at position 158 in papain, we can conclude that it is not the charge but the hydrogen-bonding interactions involving the side chain of Asp158 that contribute the most to the stabilization of the thiolate-imidazolium ion pair in papain. However, the charge and the hydrogen bonds of Asp158 both contribute to the intrinsic activity of the enzyme.     

Basic nuclear proteins of the histone-less eukaryote Crypthecodinium cohnii (Pyrrhophyta): two-dimensional electrophoresis and DNA-binding properties

Unlike typical eukaryotes, the Dinoflagellate Crypthecodinium cohnii does not contain histones but six major basic, low molecular weight nuclear proteins which represent only 10% of the DNA mass and differ from histones in their electrophoretic and DNA-binding properties. These proteins are resolved in two-dimensional electrophoresis (AUT-PAGE x SDS-PAGE). Three proteins with an apparent molecular mass of 16, 16.5 and 17 kDa (p16, p16.5 and p17) are present in addition to the major 14 kDa basic nuclear component (HCc). HCc itself is resolved in three proteins (alpha, beta and gamma). When the proteins are not reduced with 2-mercaptoethanol before 2D-PAGE, the migration of HCc alpha, beta and gamma is modified in a way which suggests the formation of both inter- and intramolecular disulfide bridges and thus, the presence of at least two cysteines. The amino-acid analysis of HCc proteins resolved in 2D gels confirms that they are lysine-rich. HCc alpha, beta and gamma as well as p16, p16.5 and p17 are removed from isolated chromatin with 0.6 M NaCl, indicating that their affinity for DNA in vivo is lower than that of core histones. Furthermore, in vitro, they bind more tightly to single-stranded than to double-stranded DNA.     

Climatic and anthropogenic effects on the sedimentation and geochemistry of Lakes Bourget,Annecy and Léman

During the last glaciation, the Rhone glacier extended as far as the region of Lyon and covered Lakes Bourget, Annecy and Léman. Glacial retreat successively freed Lakes Bourget and Annecy, it reached Geneva around 14 000 B.P. and the head of Lake Léman at about 12 000 B.P. Deposits situated between Nyon and the foot of the Jura provide a complete palynological zonation and serve as a type section for the lake core studies. A palynological horizon (about 1 800 A.D.) has been found in Lake Bourget and wood has been dated at 3 230 ± 65 B.P. (¹⁴C) in a core from Lake Léman. Three 6 m cores from each lake were sampled together with many 0.5 m cores from Léman. The short Lake Léman cores have been dated by ¹³⁷Cs and provide a precise indication of changes in the basin during recent decades. Holocene climatic variations are evident in the Léman cores and are reflected by concentrations of carbonate and organic carbon which increase at the end of the Alleröd and reach a maximum in the Atlantic Period (climatic optimum). Human settlement on the shores of Lake Annecy is shown by increases in heavy metals; increased sedimentation rates suggest settlement at the beginning of the Christian era. All the cores show increases in the organic matter and nutrients; from the turn of the century in the Léman, and later for the other lakes where eutrophication started only a few decades ago. NAI-P shows a very recent increase, around 1965 in the Leman. Heavy metals (Hg, Cd and Pb) show increases at about 30 years ago in Annecy, at about the turn of the century in Lake Léman and in the mid-19th century in Lake Bourget.

---

Résumé Lors de la dernière glaciation, le glacier du Rhône a atteint la région de Lyon et recouvert les lacs du Bourget, d'Annecy et le Léman. Son retrait a successivement libéré les lacs du Bourget, d'Annecy, puis le Léman de leur carapace de glace. Aux environs de 14 000 ans B.P. son front se trouvait au voisinage de Genéve, puis vers 12 000 B.P. à l'embouchure du Rhône en amont du Léman.La chronostratigraphie des grandes carottes (6 m) prélevées dans les trois lacs a été établie par la palynologie, sur la base des zonations obtenues dans un profil complet provenant d'un marais situé entre Nyon et le pied du Jura. Elles ont pu être contrôlées par un marqueur palynologique dans le lac du Bourget (150 B.P.) et la datation au ¹⁴C d'un fragment de bois dans le Léman (3 230 ± 65 B.P.). Une série de carottes de grand diamètre provenant de ce dernier lac, datés par le ¹³⁷Cs, ont donné une image très précise du l'évolution du milieu durant les dernières décades. D'une facon générale, toutes les carottes montrent une augmentation des teneurs en matière organique, puis en nutrients, vers le fin du 19ème siècle pour le Léman et quelques dizaines d'années plus tard pour les lacs du Bourget et d'Annecy. Le début du l'etrophisation se situe beaucoup plus tard dans le Lémen avec l'accroissement des teneurs en phosphore, principalement de ses formes NAI-P et O-P, soit aux environs de 1965.