Rifampin resistance mutation of Bacillus subtilis altering the electrophoretic mobility of the beta subunit of ribonucleic acid polymerase

Linn, Thomas; Losick, Richard; Sonenshein, Abraham L.

doi:10.1128/jb.122.3.1387-1390.1975

Cited by 39 publications

(18 citation statements)

References 14 publications

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“…To study this possibility, we searched for developmental mutants with defects in RNA polymerase. RNA polymerase mutations are easily obtained in many bacteria by selecting for resistance to the antibiotic rifampin (6,10,11,15), a specific inhibitor which is known to bind to the ,B subunit of RNA polymerase (11,14). We found that a significant class of these rifampin-resistant mutants in M. xanthus were, in fact, defective in fruiting-body formation; in at least 11 of these mutants the two phenotypes are probably caused by a single mutation.…”

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confidence: 85%

Rifampin-resistant mutants of Myxococcus xanthus defective in development

Rudd¹,

Zusman²

1979

J Bacteriol

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Rifampin, an antibiotic which is known to bind to and inhibit RNA polymerase, was used to probe the molecular regulation of development in Myxococcus xanthus. Rifampin-resistant mutants were screened for defects in fruiting-body formation. About 20% of the isolates in the initial screenings showed major defects in developmental aggregation or sporulation. Eleven independent mutants with wild-type growth rates and stable phenotypes were analyzed by transduction. In these strains, the rifampin-resistant and nonfruiting phenotypes showed cotransduction frequencies equal to or greater than 99.0 to 99.9%. The RNA polymerase activities were resistant to rifampin in vitro, indicating that the RNA polymerase is altered in these strains. Although their fruiting phenotypes are heterogeneous, these strains can be divided into two classes based on the level of aggregation. The results suggest that RNA polymerase plays a significant role in the regulation of development in M. xanthus since mutations which cause no apparent changes in vegetative growth result in striking defects in fruiting-body formation.Myxococcus xanthus is a gram-negative bacterium which grows vegetatively in soils on decaying organic material, on the bark of living trees, or by preying upon other microorganisms (7). When nutrients are depleted from a solid culture medium, cells aggregate to form mounds. Within the mounds, the rod-shaped vegetative cells convert to round or ovoid spores. Mounds of mature myxospores are referred to as fruiting bodies,We have recently investigated protein synthesis during aggregation and fruiting-body formation in M. xanthus and have found that many proteins showed significant changes in their amounts and patterns of synthesis during development (M. Inouye, S. Inouye, apd D. R. Zusman, Dev. Biol., in press). These results suggest a complex but precise program of gene expression. It is likely that the synthesis of at least some of these proteins is regulated at the transcriptional level. To study this possibility, we searched for developmental mutants with defects in RNA polymerase. RNA polymerase mutations are easily obtained in many bacteria by selecting for resistance to the antibiotic rifampin (6,10,11,15), a specific inhibitor which is known to bind to the ,B subunit of RNA polymerase (11,14). We found that a significant class of these rifampin-resistant mutants in M. xanthus were, in fact, defective in fruiting-body formation; in at least 11 of these mutants the two phenotypes are probably caused by a single mutation. The heterogeneous terminal phenotypes observed in these mutants suggests that transcriptional spec-ificities may be altered in several different ways during development. MATERIALS AND METHODSBacterial and bacteriophage strains. Three different strains that are "wild type" for fruiting-body formation were used: DZ2 (5), DZF1 (FB) obtained from D. Kaiser, who obtained it from M. Dworkin, and DZF6 (4). DZ1 (18), a nonfruiting strain, was used as a phage indicator. The phage strain MX4ts27htfhfrm (4) was used ...

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confidence: 85%

Rifampin-resistant mutants of Myxococcus xanthus defective in development

Rudd¹,

Zusman²

1979

J Bacteriol

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“…This change did not occur in Rif mutants blocked at stage 0 of sporulation (280,281), with the exception of one mutant examined by Sonenshein et al (280). Purification of RNA polymerase had indicated that the mutation to rifampin resistance alters the f3 subunit, which is in the core of the enzyme (38,186,187). Precisely how the alteration affects sporulation…”

Section: Mutations To Antibiotic Resistance That Alsomentioning

confidence: 99%

Genetic aspects of bacterial endospore formation.

Piggot¹,

Coote²

1976

Bacteriological Reviews

405

283

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95 cysA14 60 ery-1 25 purA 49 metC3; 21 ura-1 c 42 ura-26 s 83 ura-1; 5 metC3 ' 54 lys-l' 44 trpC2 35 trpC2 33 Iys-1; 16 trpC2 56 lys-1 25 phe-1; 10 ilvCl RI = 0.1 12 hisAI RI s 0.2 47 ura-1; 1 metC3 12 ura-26 16 ura-1; 49 metC3 49 52 148 307 24 phe-12; 1 lys-l 49 B 49 49, 143, 334 49 244 49 a Linkage with vegetative markers is expressed as % cotransfer. Linkage between spo mutations is expressed as the recombination index (RI [170]). b A, J.A. Hoch, personal communication; B, M. Young, personal communication; C, P.J. Piggot, unpublished observations. I Assignment to locus is not certain, see text. d If 94U (reference 148) is the same as 94 (reference 257), then this has the same phenotype as spoIVC mutants described by Coote (49, 50), and the mutations should be placed in the spoIVC locus.

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“…Recently we have observed that it is also linked to comK and is located between these two loci (170). mecB is closely linked by transformation to rifampin, streptolydigin, and lipiarmycin resistance alleles (61), which were previously shown to be in or near the rpoB gene (117,197). The latter encodes the ,1 subunit of RNA polymerase.…”

Section: Regulatory Genesmentioning

confidence: 99%