Genetic studies of the ribosomal proteins in Escherichia. coli. II. Altered 30s ribosomal protein component specific to spectinomycin resistant mutants

Summary Spectinomycin resistant (spc ^r) mutants were obtained by treating the cells of E. coli K12, W3637 with nitrosoguanidine. The compositions of ribosomal proteins were analyzed for six out of eleven such spc ^r-mutants with chromatography on a carboxymethyl cellulose (CMC) column. The 30s ribosomal subunit from all of the spc ^r-mutants was found to contain the altered 30-4 protein component, while no difference was detected in 50s ribosomal proteins between spc ^r and spc ^s bacteria.Abbreviations used CMC carboxymethyl cellulose - str streptomycin - spc spectinomycin     

Site-directed insertion mutagenesis with cloned fragments in Escherichia coli by P1 phage transduction

Summary A cloned gene with an insertion, which was made by introducing cat, was ligated to the cloning site of the phage gt11. P1 phage grown on cells lysogenized with the recombinant phage could transduce the mutant gene into the original site on the Escherichia coli chromosome.     

The temperature sensitive mutant 72c

Summary The spontaneous temperature sensitive mutant 72c is shown to be more tolerant to fusidic acid, but less tolerant to trimethoprim on plates at permissive temperature, than is the parental strain. The poor growth of the mutant on amino acids supplemented plates, as well as its inability to grow on broth plates at 40°, can be compensated by sublethal amounts of chloroamphenicol. Also some mutations to Rif-R or Str-R improve growth of the mutant under certain conditions.Reversion and other genetic analysis strongly suggest, that the pleiotropic behaviour of the mutant is due to a single mutation in a gene, which is designated fusB and is closely cotransducible with lip at min 14 of the E. coli chromosome. The gene order is lip-fusB-supE.     

Phenotypes represented by a mutational change in a 50s ribosomal protein component, 50-8, in Escherichia coli

Summary An erythromycin resistant (ery ^r) mutant of Escherichia coli Q13, QE107, was characterized by (1) the cross resistance of the cells to several macrolide antibiotics such as erythromycin, tylosin, spiramycin, oleandomycin and leucomycin, (2) the reduced affinity of its ribosomes to erythromycin and probably to the other macrolides mentioned above, (3) a low peptidyl transferase activity of its ribosomes and (4) an altered 50-8 protein of the 50s ribosomal subunits. These characters were always transferred together with the ery marker in the transduction experiments.Preliminary data of part of this work has been published (Tanaka, Teraoka, Tamaki, Watanabe, Osawa, Otaka and Takata, 1971).     

Genetic studies of the ribosomal proteins in Escherichia coli. 8. Mapping of ribosomal protein components by intergeneric mating experiments between Serratia marcescens and Escherichia coli

Summary Ribosomal protein compositions of Serratia marcescens and Escherichia coli K12 were analyzed by using carboxymethyl cellulose column chromatography. Nine 50S and nine 30S ribosomal proteins of E. coli K12 could be distinguished from those of S. marcescens on the chromatogram.Episomes of E. coli K12, which cover the streptomycin(str) region of the chromosome, were transferred to S. marcescens. Chromatographic analyses were made on the ribosomal proteins extracted from these hybrid strains. At least nine 30S and six 50S ribosomal proteins of E. coli-type could be detected in the ribosomes of the hybrid strains in addition to the ribosomal proteins of S. marcescens.     

Nucleotide sequence of the gene for Escherichia coli ribosomal protein S15 (rpsO)

Summary The nucleotide sequence of the ribosomal protein gene rpsO (S15) and its flanking region were determined. The amino acid sequence of S15 protein deduced from the nucleotide sequence is in good agreement with the published amino acid sequence with one exception. The nucleotide sequence shows two probable promoter sites about 100 nucleotides upstream from the initiation codon (AUG) of rpsO. Inspection of the sequence also revealed structural homology between the distal part of rpsO and the reported S15 binding region in 16S rRNA.     

Microbial interactions responsible for dissolved DNA production in a hypereutrophic pond

Concentration of dissolved DNA, microbial biomass, and consumption of bacteria by heterotrophic nanoflagellates (HNF) and ciliates were examined in a hypereutrophic pond for over 7 months to elucidate the main factors which influenced the release of dissolved DNA. Changes in concentration of dissolved DNA correlated well with both abundance of ciliates ( r = 0.788, p < 0.01) and rotifers ( r = 0.738, p < 0.01). A significant correlation was also found between dissolved DNA concentration and ciliate community ingestion rates ( r = 0.668, p <0.01). These results suggest that consumption of bacteria by ciliates is an important reason for the release of dissolved DNA. This revised version was published online in July 2006 with corrections to the Cover Date.     

Signaling via the G protein alpha subunit FGA2 is necessary for pathogenesis in Fusarium oxysporum

Cloning and disruption of fga1, the gene encoding the G protein alpha subunit FGA1 in phytopathogenic fungus Fusarium oxysporum, has been reported previously, and the fga1 disruptants showed altered colony morphology, increased heat resistance, reduced conidiation and pathogenicity. To further evaluate the role of G protein signaling in this fungus, cloning of fga2, which encodes the second Galpha protein FGA2, was performed by PCR methods. The deduced primary structure of FGA2 (355 amino acid residues) showed high identity with other Galpha proteins, which belong to class III of fungal Galpha proteins. Disruption of fga2 led to higher heat resistance, similar to the fga1 disruptants, but pathogenicity was completely lost, unlike the fga1 disruptants. Alteration of colony morphology and conidiation, which was observed in the fga1 disruptants, was not observed in the fga2 disruptants. The fga1/fga2 double disruptants showed phenotypic alterations similar to the fga1 or fga2 single disruptants, but increase of heat resistance was much more pronounced than in each single disruptant.