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The phage promoter responsible for the expression of the inserted beta-galactosidase gene in bacteriophage lambda plac5.

The lac transducing phage, lambda plac5, carries a segment of the E. coli lac operon on the left side of the b2 region of the lambda phage. In the absence of additional cyclic AMP, beta-galactosidase can only be expressed from the phage promoter, and the expression of the inserted lac promoter is suppressed. This phage promoter responsible for beta-galactosidase synthesis is shown to be under the control of the cI and N gene products; however, the repressive action of the cro gene product at high multiplicity of infection is not observed although some turn off at very late time is detected. To pin down this phage promoter, results described in this communication and those described elsewhere can rule out the promoter PI, PR, P'R, and the promoter PL also looks rather unlikely. No firm identification of this phage promoter has been made, but the promoter(s) in the b2 region (the b2 promoter) is proposed. The phage promoter responsible for beta-galacrosidase synthesis is shown to be a weak promoter, requires the Q gene product or one (or more) of the late gene products for activation, and the time of expression is very late.

Bacteriophage lambda↗

Control of ribosomal RNA synthesis in Escherichia coli. V. Stimulation of rrnC gene transcription in vitro by a protein factor.

Ribosomal RNA synthesis has been investigated in an in vitro system consisting of purified E. coli RNA polymerase and phi 80d3 DNA carrying rrnC operon. rRNA comprises about 25% of the total RNA synthesized in this system under optimal conditions and is stimulated by a crude protein fraction prepared from E. coli cell extracts. The stimulation activity has been fractionated by a Sephacryl S200 column and detected as a single peak of about 50,000 daltons molecular weight. The activity specifically increases the initiation frequency of transcription of the rRNA operon on the phage DNA. The addition of ppGpp partially inhibits the stimulation activity for rRNA synthesis.

Bacterial Proteins↗

Analysis of ultraviolet light-induced suppressor mutations in the strain of Escherichia coli K-12 AB1157: an implication for molecular mechanisms of UV mutagenesis.

Genetic analysis of histidine independent (His+) revertants induced by ultraviolet light in the his-4 E. coli strain AB1157 was carried out: 93% carried ochre (UAA) suppressor mutations and 17% carried back mutations to his+ or (intragenic?) suppressors not detectably separable from his-4. Using the specialized transducing lambda psu 2int- phage, which carries supE-supB, it was determined that 87% of the ochre suppressors mapped in the supE-supB region. We were able to deduce that 56% of these affected tRNA1Gln by a CAA leads to TAA change in the tRNA gene while 31% affected tRNA2Gln by TAG- leads to TAA change. Although we were unable to deduce the base substitution of the remaining 13%, the results indicate that most of the suppressor mutations are caused by G:C to A:T transition. These results suggest that the high incidence of supE-supB region suppressor mutation in E. coli by UV would be a reflection of the general feature of UV mutagenesis; i.e. preferential induction of G:C to A:T transition in repairing nonpairing DNA lesions. AI 05371

Bacteriophage lambda↗

Factors affecting transcriptional regulation of the formate-hydrogen-lyase pathway of Escherichia coli.

The regulatory elements involved in expression of the gene (fdhF) for the selenopolypeptide of formate dehydrogenase and of a gene (or transcriptional unit) (hyd) specifically responsible for the formation of the gas-evolving hydrogenase (hydrogenase 3) in Escherichia coli were investigated. Formate (or a product of it) is required for expression of both systems since in a pyruvate-formate-lyase deficient mutant induction occurs only when formate is supplemented externally. Under this condition, formate can partially overcome repression by nitrate. The transcription of both the fdhF gene and the hydrogenase-3-encoding systems is independent of the presence of a wild-type fnr gene when formate is present, supporting the view that the Fnr effect on the formation of the formate-hydrogen-lyase pathway is indirect. Mutations blocking the synthesis of a functional molybdenum cofactor also had no major affect on fdhF and hyd expression. The nucleotide sequence of the 5' flanking region of the fdhF gene was determined and the transcription start point of the fdhF gene was localized by nuclease S1 mapping. Nuclease Bal31 generated deletion clones were constructed and the regulation of their expression was studied. Anaerobic expression and induction by formate depended on the presence of a stretch of approximately 185 nucleotides upstream of the translation start. Elements mediating formate induction and oxygen or nitrate repression could not be separated physically. The regulatory features of the fdhF upstream region bear striking resemblance to systems whose expression are dependent upon upstream activating elements.

Aldehyde Oxidoreductases↗

A new host gene (groPC) necessary for lambda DNA replication.

The isolation of a bacterial mutation in a gene, designated groPC, which affects the growth of phages lambda and P2 is described. Lambda replication is severely limited in the strain, and some lambda pi mutations, which map in (or near) the P gene, allow growth. The gro mutation, groPC259, is recessive to wild type and maps between threonine (thr) and diaminopimelate (dapB) on the E. coli chromosome. The possibility that the groPC gene is concerned with host DNA replication is discussed.

Chromosome Mapping↗

Genetic analysis of rec E activities in Bacillus subtilis.

A recE mutant (recE6) of Bacillus subtilis was constructed by insertion of a selectable marker into the recE coding region. The insertional inactivation of the recE gene renders cells very sensitive to DNA damaging agents and severely impairs intermolecular recombination, but does not markedly affect plasmid interstrand annealing and intramolecular recombination. The recE6 allele was then introduced into a set of DNA repair-deficient strains of B. subtilis. The removal of DNA damage by the recF, addA addB, recH, recL and recP gene products is strictly dependent on an active recE gene product (recE-dependent pathway). On the other hand, the increased sensitization to purine adducts in the uvrA42 recE6 and polA5 recE6 strains suggests that such lethal lesions may be removed either by the recE-dependent or by the recE-independent pathway.

4-Nitroquinoline-1-oxide↗