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C L Truitt

Publications and source records attributed to C L Truitt.

7 recordsLinked to original sources

Effects on growth and sporulation of inactivation of a Bacillus subtilis gene (ctc) transcribed in vitro by minor vegetative cell RNA polymerases (E-sigma 37, E-sigma 32).

The E-sigma 37-transcribed gene ctc was inactivated by a site-specific insertion into the Bacillus subtilis chromosome. The resulting mutation inhibited sporulation by 95% at elevated temperatures (48 degrees C). If the ctc- mutation is placed in a strain that carries a mutation in the closely linked but distinct spoVC gene, ctc now affects both growth and sporulation at elevated temperatures. Growth of the ctc- spoVC285 strain was transiently inhibited when exponentially growing cultures were shifted from 37 degrees C to 48 degrees C. A similar, but less pronounced "growth lag", was also seen in a B. subtilis strain carrying only the spoVC-285 mutation. This finding suggests that both the ctc and spoVC products function in vegetatively growing B. subtilis.

Bacillus subtilis↗

Isolation of Bacillus subtilis mutants altered in expression of a gene transcribed in vitro by a minor form of RNA polymerase (E-sigma 37).

To develop a technique for identifying Bacillus subtilis genes whose products affect transcription from promoters recognized by sigma 37-containing RNA polymerase (E-sigma 37), we cloned the promoter region of a gene (ctc) that is actively transcribed in vitro by E-sigma 37 into a plasmid (pPL603B) so that a transcriptional fusion was created between ctc and a plasmid-borne chloramphenicol acetyltransferase (CAT) gene. CAT levels in B. subtilis carrying the ctc/CAT fusion plasmid varied in a manner that was consistent with the known pattern of ctc RNA synthesis. Mutagenesis of cells harboring the ctc/CAT plasmid led to the isolation of bacterial clones which displayed altered chloramphenicol resistance. Analysis of the mutants demonstrated that CAT activity was substantially changed in the mutant cells. Several of the B. subtilis mutants, both CAT overproducers and underproducers, also had acquired a sporulation-deficient phenotype. The mutations responsible for altered CAT expression were not carried on the plasmid. Analysis of RNA synthesized by mutant cells indicates that at least a portion of the mutants may be altered in the level of transcription from the ctc promoter and, hence, are likely to define B. subtilis genes which influence this process.

Acetyltransferases↗

Inheritance of extrachromosomal rDNA in Physarum polycephalum.

In the acellular slime mold Physarum polycephalum, the several hundred genes coding for rRNA are located on linear extrachromosomal DNA molecules of a discrete size, 60 kilobases. Each molecule contains two genes that are arranged in a palindromic fashion and separated by a central spacer region. We investigated how rDNA is inherited after meiosis. Two Physarum amoebal strains, each with an rDNA recognizable by its restriction endonuclease cleavage pattern, were mated, the resulting diploid plasmodium was induced to sporulate, and haploid progeny clones were isolated from the germinated spores. The type of rDNA in each was analyzed by blotting hybridization, with cloned rDNA sequences used as probes. This analysis showed that rDNA was inherited in an all-or-nothing fashion; that is, progeny clones contained one or the other parental rDNA type, but not both. However, the rDNA did not segregate in a simple Mendelian way; one rDNA type was inherited more frequently than the other. The same rDNA type was also in excess in the diploid plasmodium before meiosis, and the relative proportions of the two rDNAs changed after continued plasmodial growth. The proportion of the two rDNA types in the population of progeny clones reflected the proportion in the parent plasmodium before meoisis. The rDNAs in many of the progeny clones contained specific deletions of some of the inverted repeat sequences at the central palindromic symmetry axis. To explain the pattern of inheritance of Physarum rDNA, we postulate that a single copy of rDNA is inserted into each spore or is selectively replicated after meiosis.

Extrachromosomal Inheritance↗

Peptide maps of regulatory subunits of Bacillus subtilis RNA polymerase.

Peptide maps of four regulatory subunits of Bacillus subtilis RNA polymerase were obtained. Three sigma-like proteins (sigma 55, sigma 37, and sigma 29) as well as the transcription modification factor, delta (delta) protein, were shown to give unique peptide patterns. This observation demonstrates that each is a distinct protein species; none is derived from another by a simple proteolytic modification.

Bacillus subtilis↗

Bacteriophage lambda mutants (lambdatp) that overproduce repressor.

Lambda tp mutants, selected for their ability to form turbid plaques on lon hosts, overproduce repressor. The tp1 and tp2 mutations have been located within (or adjacent to) the cIII gene. The tp1 mutation reduced late gene expression, as measured by endolysin synthesis (in the absence of functional cI repressor) and progeny phage yield. The tp4 mutation was mapped in the cY-cII region, and complementation tests indicated that tp4 affects the diffusible product of the cII gene. The tp4 mutation also reduced progeny production, but did not markedly affect endolysin synthesis.

Coliphages↗