Natural Read-through at the UGA Termination Signal of Qβ Coat Protein Cistron

GENETIC analysis of conditional lethal mutants of Qβ, a small RNA-containing bacteriophage of Escherichia coli, has demonstrated the existence of three cistrons¹. These have been correlated² with three of the four phage-specific proteins observed after Qβ infection^2–4: gene 1 codes for the phage-specific subunit of Qβ replicase, gene 2 for the maturation protein and gene 3 for the major coat protein. Using SDS-polyacrylamide gel electrophoresis to characterize these polypeptides, Garwes et al. introduced the nomenclature I, IIa and III for the phage-specific proteins³; the corresponding molecular weights are 64,000, 41,000 and 14,000.     

Ribosomal RNA Turnover in Contact Inhibited Cells

CONTACT inhibition of animal cell growth is accompanied by a decreased rate of incorporation of nucleosides into RNA^1–3. Contact inhibited cells, however, transport exogenously-supplied nucleosides more slowly than do rapidly growing cells^4,5, suggesting that the rate of incorporation of isotopically labelled precursors into total cellular RNA may be a poor measure of the absolute rate of RNA synthesis by these cells. Recently, Emerson⁶ determined the actual rates of synthesis of ribosomal RNA (rRNA) and of the rapidly labelled heterogeneous species (HnRNA) by labelling with ³H-adenosine and measuring both the specific activity of the ATP pool and the rate of incorporation of isotope into the various RNA species. He concluded that contact inhibited cells synthesize ribosomal precursor RNA two to four times more slowly than do rapidly growing cells, but that there is little if any reduction in the instantaneous rate of synthesis of HnRNA by the non-growing cells. We have independently reached the same conclusion from simultaneous measurements on the specific radioactivity of the UTP pool and the rate of ³H-uridine incorporation into RNAs (unpublished work of Edlin and myself). However, although synthesis of the 45S precursor to ribosomal RNA is reduced two to four times in contact inhibited cells, the rate of cell multiplication and the rate of rRNA accumulation are reduced ten times. This suggests either “wastage”⁷ of newly synthesized 45S rRNA precursor, or turnover of ribosomes in contact inhibited cells Two lines of evidence suggest that “wastage” of 45S RNA does not play a significant role in this system. (1) The rate of synthesis of 45S RNA in both growing and contact inhibited cells agrees well with that expected from the observed rates of synthesis of 28S and 18S RNAs (unpublished work of Edlin and myself). Emerson has made similar calculations⁶. (2) 45S RNA labelled with a 20 min pulse of ³H-uridine is converted in the presence of actinomycin D to 28S and 18S RNAs with the same efficiency (approximately 50%) in both growing and contact inhibited cells. These results indicate that, in order to maintain a balanced complement of ribosomal RNAs, contact inhibited cells must turn over their ribosomes. We present evidence here that rRNA is stable in rapidly growing chick cells, but begins to turn over with a half-life of approximately 35–45 h as cells approach confluence and become contact inhibited.     

Functions of Invertebrate Hemoglobins with Special Reference to Adaptations to Environmental Hypoxia

The oxygen-binding properties and some associated structuralfeatures of invertebrate hemoglobins (Hb) are reviewed togetherwith the environmental conditions (particularly in stressfulhabitats) that influence oxygen-binding functions in nature.Compared to the vertebrates invertebrate Hbs display a spectacularvariation in functional properties, which is manifest withinthe major systematic groups of animals and within the differenthistological and molecular categories of the pigment. This variabilityis seen primarily to represent adaptations to the wide rangeof environmental conditions to which Hb-bearing invertebratesare subjected in nature but also to reflect certain organismicfunctions, which determine the internal conditions under whichthe Hbs work in life. These adaptations appear to be uniquelydirected towards optimizing the transfer of oxygen from theenvironmental source to the mitochondrial combustion sites.The immense structural and functional complexity of invertebrateHb systems may well compensate for a decreased organizationat the organ level compared to that in vertebrates shiftingmore of the regulatory burden to the molecular level.