Phylogenetic relationships among various helical bacteria

Helical bacteria from the generaSpirillum, Oceanospirillum, Aquaspirillum, andAzospirillum—as well asSerpens flexibilis—were characterized by oligonucleotide cataloging of 16S rRNA in order to establish their phylogenetic relationships to one another and to Gramnegative bacteria in general. The various genera of helical bacteria are not specifically related to one another (to the exclusion of nonhelical bacteria) and, where tested, the individual genera as presently constituted are not phylogenetically coherent (with the possible exception ofOceanospirillum, which may form a deep grouping).     

The intracellular distribution of adenylate kinase in the leaves of spinach,wheat and barley

Of the total adenylate-kinase activity in 10-d-old barley and wheat leaves, 40–50% is localised in the chloroplasts, while in mature spinach leaves 50–70% of the enzyme is chloroplastic. The extra-chloroplastic adenylate-kinase activity is associated with the mitochondria, very little, if any, is freely soluble in the cytoplasm. The adenylate pool of the cytoplasm could have access to adenylate-kinase activity in the intermitochondrial space because of the free permeation of adenylates across the outer mitochondrial membrane. Thus the adenylate pool of the cytoplasm could be subject to adenylate-kinase equilibrium. The mitochondrial adenylate kinase appeared to the localised exclusively in the intermembrane space.     

Asymmetric maturation of a dimeric transfer RNA precursor

Six of the eight transfer RNAs coded by bacteriophage T4 are synthesized via three dimeric precursor molecules. The sequences of two of these have been determined. Both of these precursors give rise to equimolar amounts of the cognate tRNA molecules in vivo. In contrast, even in wild-type infections, tRNA^Ile is present in ≤ 30% the amount of tRNA^Thr, with which it is processed from a common dimeric precursor.We have now determined the sequence of this dimer. In addition to the nucleotides present in tRNA^Thr and tRNA^Ile, it contains nine precursor-specific residues, located at the 5′ and 3′ termini and at the interstitial junction of the two tRNA sequences. While the three dimers share the majority of structural features in common, pre-tRNA^{Thr + Ile} is the only case in which an encoded tRNA 3′ -C-C-A terminus is present in the interstitial region.The processing of this dimer in various biosynthetic mutants has been analyzed in vivo and in vitro and shown to be anomalous in several respects. These results suggest that the apparent underproduction of tRNA^Ile can be explained by a novel processing pathway that generates a metabolically unstable tRNA^Ile product. Data from DNA sequence analysis of the T4 tRNA gene cluster (Fukada & Abelson, 1980) support the conclusion that the asymmetric maturation of this precursor is a consequence of the unique disposition of the -C-C-A sequence. These results argue that gene expression can be modulated at the level of RNA processing. The biological significance of this phenomenon is discussed in relation to evidence that tRNA^Ile has a unique physiological role.     

Late replication patterns in adult and embryonic mice carrying Searle's X-autosome translocation

Bromodeoxyuridine-dye technique analysis of X chromosome DNA synthesis in female adult and fetal mice carrying the balanced form of the T(X; 16) 16H translocation demonstrated that the structurally normal X chromosome was late replicating (and hence presumably inactive) in 93% of the adult cells and 99% of the 9-day embryo cells, with the X¹⁶ chromosome late replicating in the remaining cells. We conclude from these results that in T16H/+ females either there is preferential inactivation of the normal X chromosome or that, if inactivation is random, cell selection takes place before 9 days of development. Two 9-day female embryos with an unbalanced karyotype were also studied; both had two late-replicating chromosomes in most of their cells, one being the chromosome 16^X, the other a normal X chromosome. These results, together with the presence of a late-replicating X¹⁶ chromosome in T16H/+ adult and fetal mice, support the concept that more than one inactivation center is present on the X chromosome of the mouse because the X¹⁶ and the 16^x chromosomes can be late replicating.     

Chromosome banding in amphibia

The distribution of constitutive heterochromatin on the chromosomes of Triturus a. alpestris, T. v. vulgaris and T. h. helveticus (Amphibia, Urodela) was investigated. Sex-specific chromosomes were determined in the karyotypes of T. a. alpestris (chromosomes 4) and T. v. vulgaris (chromosomes 5). The male animals have one heteromorphic chromosome pair, of which only one homologue displays heterochromatic telomeres in the long arms; the telomeres of the other homologue are euchromatic. This chromosome pair is always homomorphic and without telomeric heterochromatin in the female animals. There is a highly reduced crossing-over frequency between the heteromorphic chromosome arms in the male meiosis of T. a. alpestris; in T. v. vulgaris no crossing-over at all occurs between the heteromorphic chromosome arms. No heteromorphisms between the homologues exist on the corresponding lampbrush chromosomes of the female meiosis. In T. h. helveticus no sex-specific heteromorphism of the constitutive heterochromatin could be determined. The male animals of this species, however, already possess a chromosome pair with a greatly reduced frequency of chiasma-formation in the long arms. The C-band patterns and the pairing configurations of the sex-specific chromosomes in the male meiosis indicate an XX/XY-type of sex-determination for the three species. A revision of the literature about experimental interspecies hybridizations, gonadic structure of haploid and polyploid animals, and sex-linked genes yielded further evidence in favor of male heterogamety. The results moreover suggest that the heterochromatinization of the Y-chromosome was the primary step in the evolution of the sex chromosomes.     

Selective lesions of acinar pancreatic cells in rats poisoned with abrin

Rats poisoned with abrin (2.5 micrograms/100 g body weight) died within 36 h with severe necrosis of acinar pancreatic cells. Incorporation in vivo of labelled amino acids into pancreatic protein was greatly impaired 6 h after poisoning. Microsomes isolated from the pancreas of poisoned rats at 6 h had a reduced capacity for protein synthesis in vitro. Incorporation in vivo of orotic acid into pancreatic RNA was decreased 12 h after poisoning.     

The characteristics of shuttling RNAs confirmed

Shuttling RNAs are recognized as molecules that migrate against steep concentration gradients from one nucleus (through the cytoplasm) into another nucleus in the same cell. In previous work these molecules were identified through experiments involving the separation of two kinds of nuclei utilizing differences in the nuclei that may have produced misleading results. In the experiments reported here normal, randomly-chosen ameba (A. proteus) nuclei containing [³²P]RNA were implanted into unlabeled normal, randomly-chosen cells and, after suitable incubation, the labeled RNAs present in each kind of nucleus were characterized by gel electrophoresis. The previously obtained results were confirmed: i.e. (a) the recipient cell nuclei acquired the same four small, distinct RNAs, which are recognized as shuttling ones because they migrate from one nucleus to the other; (b) the grafted nuclei possess, in addition to the four shuttling RNAs, three small, distinct RNAs, which are recognized as non-shuttling RNAs. New evidence also is presented to show that the acquisition by a nucleus of labeled RNAs in the above kind of experiment is not a result of new synthesis of RNAs from the labeled turnover products emanating from the transplanted nucleus.