Instability of bacteriophage Mu transposase and the role of host Hfl protein

The activity of the transposase of bacteriophage Mu is unstable, requiring the protein to be synthesized throughout the lytic cycle (Pato and Reich, 1982). Using Western blot analysis, we analysed the stability of the transposase protein during the lytic cycle and found that it, too, is unstable. The instability of the protein is observed both in the presence and the absence of Mu DNA replication, and is independent of other Mu-encoded proteins and the transposase binding sites at the Mu genome ends. Stability of the protein is enhanced in host strains mutated at the hfl locus; however, stability of the transposase activity is not enhanced in these strains, suggesting that functional inactivation of the protein is not simply a result of its proteolysis.     

Partial characterization of cell wall from a flocculent strain ofKluyveromyces marxianus

Summary Cell walls from aKluyveromyces marxianus either non flocculent or flocculent strain were isolated and analysed for protein, carbohydrates and phosphate content. Alkaline extract of proteins were analysed by SDS-PAGE. The results revealed a higher protein content in the cell walls from the flocculent strain. Electrophoresis of the cell wall proteins of the flocculent strain showed an extra peptide with an apparent molecular weight of 37 KDa which is absent fom non-flocculent cells. The involvement of this protein in cell adhesion during flocculation is discussed.     

Unusual DNA structures at the integration site of an HIV provirus

Supercoiled pHXBc2 DNA (containing the genome of the human immunodeficiency virus type 1 and human sequences) migrated more slowly than linear DNA in native and ethidium bromide agarose gel electrophoresis at 4.5 volts/cm, suggesting the presence of unusual DNA structures. S1 nuclease analysis of pHXBc2 revealed two S1 hypersensitive sites. Site I was located within a 25 bp direct repeat in host DNA 0.6 kB upstream from the 5' LTR. Site II was mapped 0.2 kB upstream from the vif gene start site. Sequence analysis showed that Site I sequences could assume different unusual DNA structures, whereas sequences at Site II could assume either slipped or H-DNA forms. Unusual DNA structures in host DNA may be associated with active chromatin regions and may favor proviral integration.     

Characterization of amber mutations in bacteriophage Mu transposase: a functional analysis of the protein

We have characterized a series of amber mutations in the A gene of bacteriophage Mu encoding the phage transposase. We tested different activities of these mutant proteins either in a sup0 strain or in different sup bacteria. In conjunction with the results described in the accompanying paper by Bétermier et al. (1989) we find that the C-terminus of the protein is not absolutely essential for global transposase function, but is essential for phage growth. Specific binding to Mu ends is defined by a more central domain. Our results also reinforce the previous findings (Bétermier et al., 1987) that more than one protein may be specified by the A gene.     

Through-bond electron-transfer interaction in proteins

A model for the through-bond electronic interaction between electron donor and acceptor in proteins is developed. We use a one-electron Hamiltonian, write the Dyson's equation in site representation and solve it by using a Green's function formalism with some renormalization ideas. An expression for Tab which describes the exponential decay with distance bond per bond is obtained. Covalent, non-covalent and convergent pathways are considered and no periodic approximation is needed.     

Localization of sequences regulating ancestral and acquired sites of esterase6 activity in Drosophila melanogaster

We have broadly defined the DNA regions regulating esterase6 activity in several life stages and tissue types of D. melanogaster using P- element-mediated transformation of constructs that contain the esterase6 coding region and deletions or substitutions in 5' or 3' flanking DNA. Hemolymph is a conserved ancestral site of EST6 activity in Drosophila and the primary sequences regulating its activity lie between -171 and -25 bp relative to the translation initiation site: deletion of these sequences decrease activity approximately 20-fold. Hemolymph activity is also modulated by four other DNA regions, three of which lie 5' and one of which lies 3' of the coding region. Of these, two have positive and two have negative effects, each of approximately twofold. Esterase6 activity is present also in two male reproductive tract tissues; the ejaculatory bulb, which is another ancestral activity site, and the ejaculatory duct, which is a recently acquired site within the melanogaster species subgroup. Activities in these tissues are at least in part independently regulated: activity in the ejaculatory bulb is conferred by sequences between -273 and -172 bp (threefold decrease when deleted), while activity in the ejaculatory duct is conferred by more distal sequences between -844 and -614 bp (fourfold decrease when deleted). The reproductive tract activity is further modulated by two additional DNA regions, one in 5' DNA (-613 to -284 bp; threefold decrease when deleted) and the other in 3' DNA (+1860 to +2731 bp; threefold decrease when deleted) that probably overlaps the adjacent esteraseP gene. Collating these data with previous studies suggests that expression of EST6 in the ancestral sites is mainly regulated by conserved proximal sequences while more variable distal sequences regulate expression in the acquired ejaculatory duct site.      

Interfacial and micellar behavior of glucose lipid

Summary Glucose lipid of notable surfactant properties was produced by using hydrocarbon assimilating bacterium of the bacterial strain MM1. Its surface active properties were notable in spite of ,-hydrophilic and bulky molecule. The critical micelle concentration (cmc) was small (0.165 M); surface and interfacial tension for hexadecane at 0.1 % (pH 7.35) solution, 24.6 and 13.3 mN/m, respectively. The emulsifying action was excellent and comparable to those of rhamnolipids. The micelle has the outer radius. 28.5 Å and inner core, 6.7 Å on the basis of the concentric spherical shell model by means of the small angle X-ray solution scattering measurement.Dedicated to Professor Fritz Wagner on the occasion of his 65th birthday.     

Phylogenetic diversification of immunoglobulin genes and the antibody repertoire

Immunoglobulins are encoded by a large multigene system that undergoes somatic rearrangement and additional genetic change during the development of immunoglobulin-producing cells. Inducible antibody and antibody-like responses are found in all vertebrates. However, immunoglobulin possessing disulfide-bonded heavy and light chains and domain-type organization has been described only in representatives of the jawed vertebrates. High degrees of nucleotide and predicted amino acid sequence identity are evident when the segmental elements that constitute the immunoglobulin gene loci in phylogenetically divergent vertebrates are compared. However, the organization of gene loci and the manner in which the independent elements recombine (and diversify) vary markedly among different taxa. One striking pattern of gene organization is the "cluster type" that appears to be restricted to the chondrichthyes (cartilaginous fishes) and limits segmental rearrangement to closely linked elements. This type of gene organization is associated with both heavy- and light-chain gene loci. In some cases, the clusters are "joined" or "partially joined" in the germ line, in effect predetermining or partially predetermining, respectively, the encoded specificities (the assumption being that these are expressed) of the individual loci. By relating the sequences of transcribed gene products to their respective germ-line genes, it is evident that, in some cases, joined-type genes are expressed. This raises a question about the existence and/or nature of allelic exclusion in these species. The extensive variation in gene organization found throughout the vertebrate species may relate directly to the role of intersegmental (V<==>D<==>J) distances in the commitment of the individual antibody-producing cell to a particular genetic specificity. Thus, the evolution of this locus, perhaps more so than that of others, may reflect the interrelationships between genetic organization and function.      

Calorific values of Chlorella vulgaris (Trebouxiophyceae) as a function of different phosphorus concentrations

Quantification of the calorific content of microalgae is critical in studies of energy flow, trophic partitioning, plant/herbivore interactions in aquaculture and biomass production for biofuels. We investigated the calorific value and biochemical composition of Chlorella vulgaris at different phosphorus (P) concentrations (6.0 × 10^?7, 2.3 × 10^?6 and 2.3 × 10^?4 mol L^?1 P). As expected, the control (2.3 × 10^?4 mol L^?1 P) supported better growth than P limited treatments. Biomolecules like total carbohydrates and lipids accumulated under P limitation, which significantly correlated with high calorific values. Lipid class composition showed that triacylglycerols were the most accumulated under P limited conditions. The calorific value reported under control conditions (13.78 kJ g^?1) was less than those obtained under P limitation (30.47–33.07 kJ g^?1). The highest calorific value with less growth retardation was obtained at 2.3 × 10^?6 mol L^?1 P.