Synthesis of Bacterial Flagella I. Requirement for Protein and Ribonucleic Acid Synthesis During Flagellar Regeneration in Bacillus subtilis

Abstract

A relatively simple immunochemical procedure for estimating flagellar protein was developed. This procedure involved measuring the binding of purified, radioactively labeled, antiflagellar antibodies to bacteria. The assay was used to determine the requirements for ribonucleic acid (RNA) and protein synthesis during flagellar regeneration in Bacillus subtilis. Immediate inhibition of flagella development was observed when chloramphenical or puromycin was added to cells. This inhibition indicated the absence of a large pool of flagella precursors that could be assembled in the absence of protein synthesis. When the cells were starved for uracil or treated with actinomycin D to inhibit RNA synthesis, the ability of the cells to regenerate flagella decayed with a half-life of 5.5 min. When B. subtilis auxotrophs were starved for tryptophan, they continued to synthesize flagella, although this process was also inhibited by actinomycin D. On the basis of these results, we concluded that (i) the system involved in flagellar regeneration does not have unusual metabolic stability, (ii) regeneration requires both concomitant protein and RNA syntheses, and (iii) B. subtilis continues to synthesize messenger RNA during tryptophan starvation.

Flagellar Synthesis inSalmonella typhimurium: Requirement for Ribonucleic Acid Synthesis

“…These are functional and physical properties characteristic of mRNA. A similar requirement for RNA synthesis during flagellar regeneration in B. subtilis has recently been demonstrated by Dimmitt et al (6).…”

Stable Messenger Ribonucleic Acid and Germination ofMyxococcus xanthusMicrocysts

“…Samples (I ml) were taken and treated as described in Materials and Methods. Symbols: 0, vegetative-cell control; A, vegetative cells plus 8 were incubated in DCT medium at 3.1 X 108 cells/ml, andL-keucine-1-14C (24.3 mc/mmole) was added at 0.05 uc/ml. Samples (2 ml) were taken and treated as described in Materials and Methods.…”

Physiology and genetics of the "ribonucleic acid control" locus in escherichia coli.

“…Radioactive RNA was extracted from cultures of RCstr and RCrel bacteria which had been pulse labeled either during exponential growth or during amino acid starvation. The results indicated that, under the two conditions, the fraction of the labeled RNA which could be characterized as mRNA by hybrid formation with DNA was essentially the same; i.e., 20 to 25% of the labeled RNA was hybridizable. Since the fraction of the labeled RNA that was characterized as mRNA seemed to be constant under all conditions, these results also support the hypothesis that the regulation of RNA synthesis is coordinate.…”

“…The conclusion that mRNA synthesis can continue during amino acid starvation was also arrived at from measurements of flagellar protein formation in B. subtilis and S. typhimurium during tryptophan starvation (20). Flagellar protein contains no tryptophan residues, so that it is possible to study its formation in cells that are tryptophan starved.…”