Thyroid membrane ADP ribosyltransferase activity. Stimulation by thyrotropin and activity in functioning and nonfunctioning rat thyroid cells in culture

Bovine thyroid membranes possess both ADP ribosyltransferase and NAD glycohydrolase activities with the same Km values for NAD and the same pH optima. In intact membranes, the ADP ribosyltransferase is limited in its extent by the amount of available membrane acceptor which can be ADP-ribosylated; in membranes solubilized with lithium diiodosalicylate, an artificial acceptor, L-arginine methyl ester, can be substituted to eliminate this limitation. The product of the ADP ribosyltransferase is a mono-ADP-ribosylated acceptor whether the intact or solubilized membrane provides the enzyme activity and whether membrane or exogenous acceptor, L-arginine methyl ester, is utilized. The intact membranes and the solubilized preparation also have an enzyme activity which can release AMP from the mono-ADP-ribosylated acceptor whether formed by the action of the membrane ADP ribosyltransferase or the A promoter of cholera toxin. The NAD glycohydrolase activity appears to represent the half-reaction of the ADP ribosyltransferase, i.e. an activity measurable substituting water for a membrane acceptor or L-arginine methyl ester. Membranes from functional rat thyroid cells in culture, i.e. cells chronically stimulated by thyrotropin and unresponsive to further additions of thyrotropin, have low ADP-ribosylation but high NAD glycohydrolase activities. In contrast, membranes from nonfunctional rat thyroid cells, i.e. cells unresponsive to thyrotropin, have high ADP-ribosylation and low NAD glycohydrolase activities. NAD hydrolysis by the NAD glycohydrolase activity cannot account for the low ADP-ribosylation activity in membranes from the functioning cells, and its low level of ADP-ribosylation can be eliminated by solubilizing the membranes and substituting an artificial acceptor, L-arginine methyl ester. The ADP ribosyltransferase activity of rat thyroid cell membrane preparations can be enhanced by thyrotropin in a dose-dependent manner but not by insulin, glucagon, hydrocortisone, adrenocorticotropin, or its glycoprotein hormone analog, human chorionic gonadotropin. It is thus suggested (i) that, in analogy to cholera toxin, thyrotropin-stimulated ADP-ribosylation may be important in the regulation of the adenylate cyclase response and (ii) that the level of membrane acceptor available for ADP-ribosylation may relate both to a stable "'activated" state of the adenylate cyclase system in cells chronically stimulated with thyrotropin and/or to a desensitized state with regard to a failure of more thyrotropin to elicit additional functional responses.     

Defective lysis of streptomycin-resistant escherichia coli cells infected with bacteriophage f2

A lysis defect was found to account for the failure of a streptomycin-resistant strain of Escherichia coli to form plaques when infected with the male-specific bacteriophage f2. The lysis defect was associated with the mutation to streptomycin resistance. Large amounts of apparently normal bacteriophage accumulated in these cells. Cell-free extracts from both the parental and mutant strains synthesized a potential lysis protein in considerable amounts in response to formaldehyde-treated f2 RNA but not in response to untreated RNA. As predicted from the nucleotide sequence of the analogous MS2 phage, the protein synthesized in vitro had the expected molecular weight and lacked glycine. The cistron for the lysis protein overlapped portions of the coat and replicase cistrons and was translated in the +1 reading frame. Initiation at the lysis protein cistron may be favored by translation errors that expose the normally masked initiation site, and streptomycin-resistant ribosomes, known to have more faithful translation properties, may be unable to efficiently synthesize the lysis protein.     

DNA sequence homology in Rhizobium meliloti plasmids

Summary Plasmids were recovered by an alkaline procedure from six symbiotically effective strains of Rhizobium meliloti of diverse geographical origin, reported to harbour only one middle-size large plasmid (ranging from 89 to 143 Megadaltons). Each purified plasmid was digested with eight restriction endonucleases; cleavage patterns were very complex: only KpnI and XbaI gave a limited number of bands. Fingerprints were very different, whatever the restriction enzyme or the geographical origin of the strains. However, Southern DNA-DNA hybridizations revealed that the plasmids showed homologous sequences having a high thermal stability. We gave evidence that some of these sequences are common to all the plasmids of R. meliloti. The biological function of these common sequences is unknown. Hybridization with cloned nitrogen fixation (nif) genes from Klebsiella pneumoniae had demonstrated that nif genes were not located on the middle — size plasmids of R. meliloti studied in this paper.     

Metabolism of propionic acid by golden delicious apples

The headspace of whole Golden Delicious apples treated with propionic acid vapour, was analysed by means of GC, after enrichment on Tenax GC, and its c     

Functional aspects of Abscisic Acid Metabolism in Cotyledons of Phaseolus vulgaris L. Seedlings

During the early stages of germination and vegetative development,cotyledons of intact bean (Phaseolus vulgaris L.) seedlingsshowed active ABA catabolism causing a low endogenous ABA content.At the end of the substrate mobilizing phase, when the cotyledonsbecame senescent, a drastic increase of the endogenous ABA contentlinked with a decrease of the ABA catabolic activity was observed.This indicates that a close connection exists between senescenceand endogenous ABA content and metabolism in bean cotyledons. Removal of the apical growth region induced in the cotyledonsactivation of the ABA catabolism and the endogenous ABA concentrationdecreased below the detection limit (0.1 ng/g fr wt) at theonset of the outgrowth of the axillary buds. From then, apicaldominance was restored and the cotyledons returned to the senescentstate, which was correlated with a drastic increase of theirendogenous ABA content. ¹ Bevoegd verklaard navorser N. F. W. O. ² Wetenschappelijk medewerker F. K. F. O. (Received November 25, 1980; Accepted February 13, 1981)     

The relation between spruce monoculture and incipient podzolisation in ochreous brown earths of the Belgian Ardennes

Summary The effects of coniferous monoculture on the distribution of C, Fe and Al in the upper horizons of brown ochreous earths of the Belgian Ardennes were investigated by comparing 5 soil profiles developed under an 80 year old spruce stand with another 5 profiles developed under a climactic broad-leaved forest (beechwood).Organic carbon, Fe and Al were extracted with 0.1N NaOH/Na-tetraborate solution buffered at pH 9.7: recent studies have shown that this extraction is particularly appropriate for the detection of incipient podzolisation in brown earths-brown podzolic soils intergrades.Even if most of the classic podzolisation indexes fail to illustrate differences, nevertheless our results show that fulvic acids and organo-ferric complexes are present in significantly greater amounts in the upper part of the cambic (B)₁ horizon of the soils developed under conifers. Moreover, this podzolic tendancy is confirmed by the weathering patterns of the clay minerals in the A₁(B) horizons developed under spruces, i.e. a more pronouced weathering of chloritic layers than those observed in the beechwood soil, with a correlative genesis of more abundant smectite-like minerals. One may therefore conclude that the change in the humus type (moder to mor) after the planting of spruce trees, has been sufficient, within the local climatic and edaphic context, to promote incipient podzolisation.     

Tunicamycin inhibition of carbohydrate incorporation into thyroglobulin

Carbohydrate chains formation into thyroglobulin (Tg) is a prerequisite for thyroid hormones formation and completeness of carbohydrates chains is necessary for secretion of Tg into the follicles. Tg biosynthesis has been investigated by in vitro experiments, incubating rat thyroid glands with labeled amino-acid and carbohydrate in the presence of tunicamycin, a specific inhibitor of protein glycosylation. Tunicamycin inhibit Tg biosynthesis which is impaired in carbohydrate chains addition but slightly in the polypeptide synthesis, as shown by inhibition of 3H-glucosamine incorporation. Thus tunicamycin inhibits carbohydrate incorporation into Tg without affecting the polypeptide chain growth and decreases its secretion into the follicles.     

Basis and extent of genetic variability among doubled haploid plants obtained by pollen culture in Nicotiana sylvestris

Summary All diploid plants (doubled-haploid plants: D.H.) regenerated by androgenesis from binucleated pollen grains in Nicotiana sylvestris differ genetically from the original line as far as morphological features and growth rates are concerned. This androgenic variation (A.V.) is under nuclear control and is transmitted continuously by some D.H. for at least four generations of selfing; other D.H. progenies segregate. Further androgeneses carried out on one single D.H. reveal a new variability and increase the drift from the original line. All results cannot be explained by the presence of residual heterozygosity in the original line, and we suggest that most of the A.V. could originate from changes that occur in the DNA of the vegetative pollen grain cell. D.H. resulting from endomitosis of the vegetative cell would be homozygous and stable, whereas D.H. resulting from nuclear fusion between a vegetative and a generative cell would be heterozygous and would segregate in seeds through succeeding generations.     

The value of electrocardiographic R-wave changes in exercise testing: Preexercise versus postexercise measurements

To determine the usefulness of R-wave amplitude changes during exercise testing for the diagnosis of coronary artery disease (CAD) and to understand the discrepancies that have been described in the literature regarding their value, we studied two groups of patients by means of electrocardiographic (EKG) treadmill testing and coronary arteriography. Group I was composed of 149 patients who were studied prospectively. The specificity of R-wave changes measured from preexercise to immediately postexercise (SRV(5)) was 81%, but that of R-wave changes measured from preexercise to peak exercise (URV(5)) was 46%. A group of 156 patients (Group II) evaluated retrospectively showed a high specificity for the SRV(5) (84%) and poor specificity for the URV(5) (39%). The sensitivity of the SRV(5) was 38% in Group I and 42% in Group II. Therefore, if measured during the immediate postexercise period and not at peak exercise, changes in R-wave amplitude may be of value in the diagnosis of coronary artery disease by electrocardiographic exercise testing.