Turnover of nascent proteins in HeLa-S3 cells and the quasi-linear incorporation kinetics of amino acids

The turnover of nascent proteins in HeLa S-3 cells was estimated simultaneously with synthesis by the difference between the expected accumulation of radioactivity, based on rate determinations from very short pulse treatments, from that observed by continuous exposure for 100 min. The results were similar to those from conventional pulse-chase analysis, but not identical for reasons which are discussed. They confirm that extensive protein turnover accompanies synthesis in the growing cell, and therefore net synthesis is much lower than gross synthesis. Cells retained all proteins synthesised during heat treatment at 45 degrees C, indicating the complete inhibition of protein degradation, cells retaining all proteins synthesised. This particular perturbation experimentally validated the deficit method, and its potential usefulness in future turnover studies.     

Sequence interrelationships of the subunits of vicilin from pea seeds

Serological studies and comparison of N-terminal amino acid sequences with the amino acid sequence deduced from a cDNA clone have been used to establish the sequence relationships between the subunits of the pea seed storage protein, vicilin. Subunits smaller than M_r～50 000 (i.e., M_r 34 000, 30 000, 25 000, 18 000, 14 000, 13 000 and 12 000) show extensive homology with molecules within M_r～50 000 group. Both the sequencing and serological data confirm earlier evidence from studies on vicilin synthesisin vivo andin vitro which indicated that the vicilin subunits smaller than M_r～50 000 arose by endoproteolytic cleavage of parent molecules within the M_r～50 000 group. Cleavage in different M_r 50 000 parent molecules containing either one or both of two susceptible processing sites accounts for the formation of all the vicilin subunits smaller than M_r～50 000, with the possible exception of the M_r34 000 polypeptide. The position of these sites in the putative parents were defined by reference to a complete amino acid sequence deduced from the sequence of DNA complementary to mRNA for one member of the M_r～50 000 group.     

Potential problems in the use of oligonucleotide probes for staphylococcal enterotoxin genes

Oligonucleotide probes unique to the five major enterotoxin genes of Staphylococcus aureus were synthesized and used to detect DNA sequences homologous to these genes in 27 non-clinical isolates of Staph. aureus isolated from nasal swabs of 74 healthy human volunteers. Genomic DNA from all 27 isolates reacted with at least one of the probes. In a phenotypic assay for toxin production by a reverse passive latex agglutination test however, only 15 of the 27 isolates produced enterotoxin in culture. The results raise the possibility that a number of Staph. aureus isolates harbour DNA sequences that are apparently silent or mutant copies of the enterotoxin genes. This complicates the identification of enterotoxin producers by tests which depend on oligonucleotide or DNA hybridization.     

Effects of Simulated Rain on the Persistence of Beauveria bassiana Conidia on Leaves of Alfalfa and Wheat

The effect of simulated rain at two intensities (26.7 or 112.7 mm h-1) and durations (30 or 60 min) on the persistence of Beauveria bassiana conidia applied in water on leaves of alfalfa leaves of alfalfa (Medicago sativa) or wheat (Triticum aestivum) was investigated. Initial populations of conidia on the leaves of both crops were similar, and ranged from 1.7 105 to 3.5 105 colony-forming units cm-2. The simulated rain reduced the concentration of conidia on leaves by 28-61%. Although there was a slight effect due to rain intensity for alfalfa, there was no influence of either rain duration or crop type on the retention of B. bassiana conidia.     

Advance of mitosis by histone phosphokinase

The previous observation that growth-associated histone kinase (HKG) from Ehrlich ascites cells brings forward mitosis in Physarum polycephalum has been confirmed with more step 1 histone kinase and a more purified (step 2) histone kinase and the statistical significance of the results assessed. The mitosis appears normal in the phase contrast microscope and DNA synthesis is initiated after mitosis as usual. In vitro the growth-associated histone kinase phosphorylates chromatin, the phosphate appearing in F 1 histone. The results are interpreted as providing support for the hypothesis that growth-associated histone kinase controls the initiation of mitosis through F 1 histone phosphorylation and chromosome condensation.