Length heterogeneity of the large spacer of Vicia faba rDNA is due to the differing number of a 325 bp repetitive sequence elements

Summary The DNA fragments including the whole large spacer region of Vicia faba rDNA were cloned in plasmid pBR325. Sixteen clones were classed into five groups which differed from each other in the lengths of the rDNA inserts. Physical maps of these length variants cloned were constructed using EcoRI, SalI, HpaI, MluI and AccI and evidence was obtained that the length heterogeneity was due mainly to the differing number of 325 base pairs (bp) subrepeating elements in the large spacer. Sequence analysis of this subrepeating element revealed that it consisted of a duplet of an approximately 155 bp sequence and a 14 bp unrelated sequence. This structure of the repetitive element is novel.     

Overlapping of the coding regions for alpha and gamma components of penicillin-binding protein 1 b in Escherichia coli

Summary The mode of biosynthesis of penicillin-binding protein(PBP)-1 b in Escherichia coli was investigated by use of the plasmid carrying the ponB(PBP-1 b) gene region. Analyses of the products synthesized in minicells and in vitro showed that PBP-1 b was synthesized as two molecular species corresponding to the and components of PBP-1 b. The coding regions for the and components were located within the ca. 3.7 kb MluI-HincII fragment and transcribed in the direction from the HincII to the MluI site. The capacity for producing the component was abolished by a deletion extending to the MluI site ca. 0.7 kb inward from the HincII end of the ca. 3.7 kb fragment; the remaining 3.0 kb region with the MluI site at both ends directed the production of the component alone. The production of the component was enough to correct all the known defects caused by a ponB mutation. In addition to these results, the analyses for cross-reacting materials produced in correspondence to the various deletions indicated that the coding regions for the and components overlapped and that the N-terminal portion was responsible for the difference between the two components. The distal region about 0.7 kb long inward from the MluI end of the MluI-HincII fragment was dispensable for producing the functional PBP-1 b, although the PBP-1 b produced was curtailed. By a larger distal deletion reaching almost to the middle of the MluI-HincII fragment, the polypeptide produced for PBP-1 b lost the ability to bind penicillin and still retained a low but significant activity for glycan synthesis. We suggest, therefore, that the polypeptide portion required for transglycosylase activity resides on the N-terminal half of PBP-1 b, followed by the middle portion necessary for penicillin-binding and the C-terminal part dispensable for the function of PBP-1 b.     

Further evidence that central neurotensin inhibits pituitary prolactin secretion by stimulating dopamine release from the hypothalamus

Intracerebroventricular (icv) injection of neurotensin (NT) (2 micrograms/rat) suppressed prolactin (PRL) release induced by L-5-hydroxytryptophan (1 mg/100 g body wt, iv), prostaglandin E2(1 microgram/rat, icv), and FK33-824 (10 micrograms/100 g body wt, iv), a Met5-enkephalin analog, in urethane-anesthetized or conscious rats. In contrast, NT did not suppress elevated plasma PRL levels sustained by a large dose of domperidone (10 micrograms/100 g body wt, iv), a peripheral dopamine antagonist. In in vitro experiments, NT (10(-5) M) stimulated dopamine release from perifused rat hypothalamic fragments. These results suggest that central NT inhibits PRL secretion by stimulating dopamine release from the hypothalamus into hypophysical portal blood in the rat.     

The modulating influence of the fluidity of cell membrane on excision repair of DNA in UV-irradiated Escherichia coli

When the extent of liquid holding recovery (LHR) was measured as a function of the temperature at the time of liquid holding and the Arrhenius plot was made, two distinctive phases for the LHR were demonstrated in UV-irradiated RecA- derivative of E. coli ole28E1, which are unable to synthesize and degrade unsaturated fatty acids. The inflection temperatures were 17-18 degrees C, 23-24 degrees C and 28-30 degrees C for linoleate-, oleate- and elaidate-grown cells, respectively. These temperatures well corresponded to the phase transition temperatures of the cell membrane supplemented with the fatty acid. It is therefore concluded that at least a component involved in in vivo excision repair in E. coli is associated with cell membrane.     

Excretion of the penicillinase of an alkalophilic Bacillus sp. through the Escherichia coli outer membrane.

Two plasmids containing the penicillinase gene of alkalophilic Bacillus sp. strain 170, pEAP1 and pEAP2, were constructed. Most of the penicillinase produced by Escherichia coli, which carried these plasmids, was found in the culture medium. This excretion is caused by the cloned DNA fragment which contains some component that changes the outer membrane of E. coli.     

Purification and properties of rat brain dipeptidyl aminopeptidase

Dipeptidyl aminopeptidase, which hydrolyzes the 7-(Gly-Pro)-4-methylcoumarinamide, has been purified from the brains of 3 week-old rats. It was purified about 2,600-fold by column chromatography on CM-cellulose, hydroxyapatite and Gly-Pro AH-Sepharose. This enzyme hydrolyzed Lys-Ala-beta-naphthylamide well with an optimum pH of 5.5. It was inhibited by diisopropyl fluorophosphate, phenyl-methanesulfonyl fluoride, some cations, and puromycin, but was not inhibited by p-chloromercuribenzoate, N-ethylmaleimide, dithiothreitol, EDTA, iodoacetic acid, and bacitracin, indicating that rat brain dipeptidyl aminopeptidase is a serine protease. This enzyme showed a molecular weight of 220,000 by gel filtration and of 51,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The properties of purified rat brain dipeptidyl aminopeptidase were similar to those of bovine pituitary dipeptidyl peptidase II, but the molecular weight and substrate specificity of these enzymes were different.     

Enhancement of adipose S-100 protein release by catecholamines

When rat epididymal fat-pad pieces were incubated in vitro with 10 microM epinephrine, S-100 protein in the tissue was markedly decreased by release into the medium. The release of adipose S-100 protein was also enhanced by norepinephrine (10 microM), isoproterenol (10 microM), and dibutyryl cyclic AMP (5 mM), but not by insulin (0.8 microM). The enhancement of S-100 protein release by 10 microM epinephrine was completely inhibited by 10 microM propranolol. These results suggest that the release of adipose S-100 protein is regulated by the beta-adrenergic effect of catecholamines.     

Sex difference in the paradoxical response of serum GH to thyrotrophin releasing hormone in cancer patients

It was demonstrated that thyrotropin-releasing hormone (TRH) elicited a paradoxical increase in basal GH levels in cancer patients. Out of 94 cancer patients, 50 were found to be GH responders and this phenomenon was more frequently recognized in female than in male cancer patients. In cancer patients under 59 years of age, the GH response to TRH was significantly greater in females than in males, although there was no sex difference in the GH response in patients above 60 years of age. In female cancer patients, the GH response to TRH was significantly greater in patients under 59 years of age than in patients above 60 years of age, while there was no age difference in the GH response in male cancer patients. It was concluded that paradoxical responses of serum GH to TRH were recognized in 53 per cent of cancer patients and were more frequently observed in female than in male cancer patients.