Characterization of a new class of circular DNA molecules in yeast

This paper describes the isolation of a pure population of covalently closed circular twisted DNA molecules from yeast. These molecules are homogeneous in size, that is consist of monomers of 2.2μ and of multiple length oligomers of n x 2.2μ. While no data rule out the mitochondrial origin of this DNA, its actual intracellular localization remains unknown; it displays the same buoyant density as the main nuclear DNA and therefore is not the heavy nuclear satellite DNA (γ-DNA described by Moustacchi and Williamson (1966)); although circular molecules represent only 1 to 5 % of the total DNA, they can be prepared in sizable and reproducible amounts by a method based on the use of mechanical disruption of yeast cells rather than lysis by snail gut juice.     

Cellular Studies of X-Ray Induced Inhibition of Lens Regeneration

Whole-body X-irradiation of adult newts 0 to 3 days after lentectomy inhibits transformation of the dorsal iris epithelium into a lens in all cases. The first question raised was whether irradiation affects infiltration of the iris area by macrophages, and the phagocytic activities of these cell types in the iris epithelium (prominent phenomena in this system). The number of macrophages infiltrating into the iris epithelium, and their phagocytic activities (indicated by uptake of melanosomes) were not affected by irradiation under those conditions. The second group of experiments concerns the possible effects of irradiation on DNA replication of iris epithelial cells, which become transformed into lens cells in the non-irradiated system. Autoradiographic studies of iris epithelial cells in vivo revealed a significant suppressive effect of irradiation on the frequencies of cells incorporating ³H-thymidine 7 and 14 days after lentectomy. When autoradiography was applied to the primary pure culture of iris epithelial cells at different time intervals after the start of culture and irradiation in vitro , significant and persistent reduction of cell labelling due to irradiation, was demonstrated. Multiplication of spread cells in the iris epithelial culture was strongly and persistently inhibited throughout a period of 2 months. Inhibition of cell labelling and of cell multiplication was always accompanied by reduction in the extent of de-pigmentation of iris epithelial cells. De-pigmentation is one of the requirements for the cells become transformed into lens cells. The possible mechanism of radiation-induced inhibition of lens regeneration is discussed.     

Differential Tolerance of Streptomycetes to Sodium Chloride as a Taxonomic Aid

A survey was made of the NaCl tolerance of approximately 1,300 Streptomyces strains belonging to 313 species. The growth medium of the organisms was supplemented with a graded series of NaCl concentrations (4, 7, 10, and 13%). Only 1.8% of the species could not tolerate 4% NaCl; 26.9% could grow at a maximum of 4%; 49.7% could tolerate a maximum of 7%; 18.8% could grow at a maximum of 10%; and only 2.8% could tolerate 13% NaCl. In evaluating the relationships of NaCl tolerance to various taxonomic features, higher tolerance was statistically associated with the "yellow" and possibly the "white"-spored streptomycetes, whereas the "red"-spored series tended to have lesser tolerance. Higher tolerance was also indicated for spiny-spored species, as a group, than for smooth-spored forms. Likewise, nonproducers of melanin, collectively, were more NaCl tolerant than melanin-producing species. Uniformity of test responses between strains of species studied suggested the usefulness of NaCl tolerance as a taxonomic criterion.     

Modelling of the three-dimensional architecture of group I catalytic introns based on comparative sequence analysis

Alignment of the 87 available sequences of group I self-splicing introns reveals numerous instances of covariation between distant sites. Some of these covariations cannot be ascribed to historical coincidences or the known secondary structure of group I introns, and are, therefore, best explained as reflecting tertiary contacts. With the help of stereochemical modelling, we have taken advantage of these novel interactions to derive a three-dimensional model of the conserved core of group I introns. Two noteworthy features of that model are its extreme compactness and the fact that all of the most evolutionarily conserved residues happen to converge around the two helices that constitute the substrate of the core ribozyme and the site that binds the guanosine cofactor necessary for self-splicing. Specific functional implications are discussed, both with regard to the way the substrate helices are recognized by the core and possible rearrangements of the introns during the self-splicing process. Concerning potential long-range interactions, emphasis is put on the possible recognition of two consecutive purines in the minor groove of a helix by a GAAA or related terminal loop.     

Signalling by protein kinase C isoforms in the heart

Understanding transmembrane signalling process is one of the major challenge of the decade. In most tissues, since Fisher and Krebs's discovery in the 1950's, protein phosphorylation has been widely recognized as a key event of this cellular function. Indeed, binding of hormones or neurotransmitters to specific membrane receptors leads to the generation of cytosoluble second messengers which in turn activate a specific protein kinase. Numerous protein kinases have been so far identified and roughly classified into two groups, namely serine/threonine and tyrosine kinases on the basis of the target amino acid although some more recently discovered kinases like MEK (or MAP kinase kinase) phosphorylate both serine and tyrosine residues.Protein kinase C is a serine/threonine kinase that was first described by Takai et al. [1] as a Ca- and phospholipid-dependent protein kinase. Later on, Kuo et al. [2] found that PKC was expressed in most tissues including the heart. The field of investigation became more complicated when it was found that the kinase is not a single molecular entity and that several isoforms exist. At present, 12 PKC isoforms and other PKC-related kinases [3] were identified in mammalian tissues. These are classified into three groups. (1) the Ca-activated -, -,and -PKCs which display a Ca-binding site (C2); (2) the Ca-insensitive -, -, -, -, and -PKCs. The kinases that belong to both of these groups display two cystein-rich domains (C1) which bind phorbol esters (for recent review on PKC structure, see [4]). (3) The third group was named atypical PKCs and include , , and -PKCs that lack both the C2 and one cystein-rich domain. Consequently, these isoforms are Ca-insensitive and cannot be activated by phorbol esters [5]. In the heart. evidence that multiple PKC isoforms exist was first provided by Kosaka et al. [6] who identified by chromatography at least two PKC-related isoenzymes. Numerous studies were thus devoted to the biochemical characterization of these isoenzymes (see [7] for review on cardiac PKCs) as well as to the identification of their substrates.This overview aims at updating the present knowledge on the expression, activation and functions of PKC isoforms in cardiac cells. (Mol Cell Biochem 157: 65–72, 1996)