Secretion of trans-zeatin by Agrobacterium tumefaciens: A function determined by the nopaline Ti plasmid

Production of the plant cytokinin, trans-zeatin, by a number of strains of Agrobacterium tumefaciens was measured by a combination of traceenrichment, HPLC and radioimmunoassay and confirmed by mass spectrometry. Secretion of trans-zeatin into a culture medium is a constitutive function of those strains which harbor a nopaline Ti plasmid. Strains cured of the nopaline Ti plasmid and those which harbor octopine or agropine plasmids are non-producers. Reacquisition of nopaline plasmids by cured strains restores production.     

Safety evaluation of nuclear polyhedrosis virus replication in pigs

To evaluate the hygienic risk involved in using baculoviruses for insect pest control, safety studies are required. Pigs were chosen as representative test animals of commercial and agricultural importance. The tests were aimed at detecting virus propagation, immune reactions, and signs of acute infection (changes in body temperature and hematology profile, swelling of lymph nodes). Four of five animals inoculated with nuclear polyhedrosis virus showed a slight temperature rise at day 2 postinfection. After day 4 postinfection, no differences between infected animals and controls were observed. In the bioassay, virus activity could be recovered from fecal samples; however, no activity was found in organ extracts. The data did not indicate hygienic risks involved in the application of nuclear polyhedrosis virus, especially that from Mamestra brassicae, in biological pest control.     

Amino acid sequence in the reactive site region of an acid-stable inhibitor of trypsin, chymotrypsin and intracellular proteinases from rabbit serum

The thermo- and acid-stable trypsin, chymotrypsin and intracellular proteinases inhibitor (TAS-inhibitor) from rabbit serum was digested by trypsin, and its domain (Mr 6200) with antitryptic activity was obtained in homogeneous state. The N-terminal amino acid sequence of this domain was established by automatic Edman degradation: Thr-Val-Ala-Ala-Cys-Asx-Leu-Pro-Ile-Val-Pro-Gly-Pro-X-Arg-Gly-Ile-Phe-X- Leu-X-Ala-Phe-X-Ala-Val-X-Gly. A high degree of homology of the primary structures rabbit, human and bovine TAS-inhibitors was demonstrated.     

RNA polymerase-rifamycin. A molecular model of inhibition

By fluorimetric titration of Rifs (E. coli B) and Rifr (E. coli rpoB255) RNA polymerases with rifamycin, the mutant polymerase was demonstrated to bind rifamycin. A comparison of spatial structures of rifamycin and dinucleotide fragment of RNA in the hybrid with DNA revealed their similarity. Taking into account this structural similarity and also the fact that two phosphodiester bonds can be formed by RNA polymerase in the presence of rifamycin, a model for the inhibition mode was proposed. According to this model, rifamycin occupies the place of two terminal nucleotides of synthesized, but not translocated pentanucleotide in the transcribing complex. Asp-516 of the wild type beta-subunit was assumed to form a hydrogen bond with the rifamycin C(23) hydroxyl group. On the base of this model, reduced "cycling" synthesis of tetra-, penta-... up to decanucleotides by the Rifr RNA polymerase, in comparison with Rifs, was predicted.     

Increased cotransformation of distant markers and altered patterns of DNA-cell interactions following the exposure of transforming DNA to two carcinogenic and mutagenic alkylating agents,diethyl and dimethyl sulfate

Transforming DNA exposed to either diethyl sulfate (diES) or dimethyl sulfate (diMS) is inactivated. The rate of inactivation depends on the marker tested and on the chemical used: diMS is more active than diES. Cotransformation of linked markers is similarly depressed. In contrast, there is a transient increase in the cotransformation of distant, unlinked markers. These observations indicate that some of the intermolecular complexes of transforming DNA created in the test tube by the treatment with diES and diMS are biologically active. Radioactively labeled DNA treated with diES or diMS changes its patterns of interaction with cellular surfaces that are characteristic of untreated DNA. A possibility is considered that such alterations in DNA-protein interactions as well as the ability of these alkylating agents to transpose fragments of chromosomal material may play an important role in the processes of mutagenesis and, especially, carcinogenesis.     

Lymphocyte activation and phospholipid pathways. 31P magnetic resonance studies

31P NMR spectra of perfused lymphocytes, embedded in alginate capsules and activated by interleukin-2, were remarkably different from those of control lymphocytes. The main differences were the appearance and gradual increase in phosphodiester signals, glycerophosphocholine and glycerophosphoethanolamine. These metabolic changes also occurred following perfusion with phorbol ester and after incubation with phytohemagglutinin (PHA) and were not dependent on a special growth medium. Nifedipine, a calcium channel blocking drug, inhibited the effects of phytohemagglutinin, but not of interleukin-2. There were no NMR spectral differences between peripheral lymphocytes, stimulated for 3 weeks, and tumor-infiltrating lymphocytes. Thus, sustained accelerated turnover of phosphatidylcholine and phosphatidylethanolamine is an inherent feature of the activation process. 31P NMR spectra of lymphocytes are characterized by a low signal of phosphocholine. Perfusion studies with high concentrations of choline and the use of dapsone, an inhibitor of cytidylyltransferase, indicated that choline kinase plays a key role in regulating phosphaditylcholine synthesis in human lymphocytes.     

Two doses of the paternal Tme gene do not compensate the lethality of the Thp deletion

The hairpin-tail (Thp) deletion in chromosome 17 is lethal when it is inherited from the mother, whereas heterozygotes with Thp deletion that is paternal in origin are viable. The lethal effect of maternal Thp is due to a deficiency of the Tme gene that is located in the Thp-deleted region. In this article we describe analysis of the viability of mice with tertiary trisomy of chromosome 17, Ts(17(16]43H, with different doses of the paternal and maternal Tme alleles. We demonstrate that the presence of an additional copy of the region with the Tme gene in the female gamete entirely compensates maternal Thp lethality. We failed to compensate the absence of the Tme gene from the chromosome of maternal derivation by two doses of Tme derived from the father. Thus evidence was obtained indicating that there are significant differences between the activities of the paternal and maternal alleles of the Tme gene due to chromosome imprinting.     

Carboxypeptidase yscS: gene structure and function of the vacuolar enzyme

The gene encoding carboxypeptidase yscS in Saccharomyces cerevisiae, CPS1, was cloned by complementation of the cps1-3 mutation. The cloned CPS1 gene, which again enabled a leucine auxotrophic cps1-3 mutant to grow on the modified dipeptide Cbz-Gly-Leu (Cbz, benzyloxycarbonyl) as sole leucine source, was sequenced and found to consist of an open reading frame of 1728 bp encoding a protein of 576 amino acids. The putative protein contains a hydrophobic stretch of 20 amino acids and a putative signal sequence cleavage site. Five putative N-glycosylation sites are also in the protein sequence. This data is consistent with the previous finding of carboxypeptidase yscS being a vacuolar peptidase. Chromosomal disruption of the CPS1 gene completely abolishes carboxypeptidase yscS activity. This protein is yet another member of the peptidases in S. cerevisiae involved in nitrogen metabolism.