RNA-POLYMERASE (RPOB) MUTANTS SELECTED FOR INCREASED RESISTANCE TO GYRASE INHIBITORS IN SALMONELLA-TYPHIMURIUM

被引：17

作者：

BLANCPOTARD, AB ^{[1
]}

GARI, E ^{[1
]}

SPIRITO, F ^{[1
]}

FIGUEROABOSSI, N ^{[1
]}

BOSSI, L ^{[1
]}

机构：

[1] UNIV PARIS 06, CNRS, CTR MOLEC GENET, F-91198 GIF SUR YVETTE, FRANCE

来源：

MOLECULAR AND GENERAL GENETICS | 1995年 / 247卷 / 06期

关键词：

GYRASE INHIBITORS; RPOB; SUPERCOILING; TRANSCRIPTION;

D O I：

10.1007/BF00290399

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Some rifampicin-resistance (Rif(R)) mutations make bacteria slightly resistant to the gyrase inhibitors novobiocin (Nov) and nalidixic acid (Nal). This suggested that it might be possible to isolate rpoB mutants using either drug for positive selection. In an inital test, we confirmed the presence of Rif-resistant isolates among clones selected for Nov resistance. These mutants are also more resistant to Nal. In a subsequent experiment, we found that mutants selected for low-level resistance to Nal include isolates harboring mutations genetically linked to the rpoB locus; of two such mutants studied, one is temperature-sensitive for growth. These two mutants, which are only marginally affected in their response to Nov, are normally sensitive to Rif and thus might be representative of a new class of rpoB alleles. The Rif-resistant and Rif-sensitive rpoB alleles that increase resistance to gyrase inhibitors have one property in common: they all suppress, to varying degrees, the defect in his operon regulation (transcriptional deattenuation) caused by a gyrase defect or inhibition by novobiocin. To further analyse the transcription-supercoiling relationships in these mutants, we examined the ability of RNA polymerase to recruit gyrase activity during transcription. This was done by two independent approaches: (i) observing transcription-induced accumulation of hyper-negatively supercoiled plasmid DNA in a topA mutant background and (ii) measuring transcription-induced plasmid DNA cleavage in the presence of oxolinic acid. Results indicate that the rpoB alleles described in this study diminish the recruitment of gyrase activity by the transcription process. This property correlates with a decrease in the rate of transcription initiation. We discuss how such a defect might account for the changes observed in gyrase-related phenotypes.

引用

页码：680 / 692

页数：13

共 73 条

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