Osmotic regulation of the streptomyces lividans thiostrepton-inducible promoter, ptipA

Nasima Ali, Paul R. Herron, Meirwyn C. Evans, Paul J. Dyson

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Transcriptional activation of the thiostrepton-inducible promoter, ptipA, in Streptomyces lividans is mediated by TipAL. This transcriptional activator belongs to the MerR/SoxR family that characteristically binds an operator sequence located between the -10 and -35 hexamers normally occupied by RNA polymerase. As for the Escherichia coli merT promoter, the ptipA hexamers are separated by a long 19 bp spacer and hence a topological transition of the DNA is likely to be a requisite for alignment with RNA polymerase. Growth conditions that could facilitate this conformational change were investigated using transcriptional fusions of ptipA with reporter genes. Adjustment of growth medium osmolarity led to increased and prolonged TipAL-dependent expression, both with and without the inducer, thiostrepton. These effects correlated with increases in negative DNA supercoiling. Moreover, an inability to induce the promoter with thiostrepton in strain TK64 was corrected by increasing the concentration of osmolyte, compensating for an apparent reduced level of negative DNA supercoiling in the strain. Prolonging the time of activation of tipA in the wild-type by manipulating growth conditions revealed that mycelial autolysis could be induced by thiostrepton in 4-d-old cultures.
LanguageEnglish
Pages381-390
Number of pages9
JournalMicrobiology
Volume148
Issue number2
Publication statusPublished - Feb 2002

Fingerprint

Thiostrepton
Streptomyces lividans
DNA-Directed RNA Polymerases
DNA
Growth
Autolysis
Reporter Genes
Osmolar Concentration
Transcriptional Activation
Escherichia coli

Keywords

  • streptomyces
  • gene expression
  • morphological development
  • DNA supercoiling
  • disease
  • immunology
  • microbiology

Cite this

Ali, Nasima ; Herron, Paul R. ; Evans, Meirwyn C. ; Dyson, Paul J. / Osmotic regulation of the streptomyces lividans thiostrepton-inducible promoter, ptipA. In: Microbiology. 2002 ; Vol. 148, No. 2. pp. 381-390.
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Osmotic regulation of the streptomyces lividans thiostrepton-inducible promoter, ptipA. / Ali, Nasima; Herron, Paul R.; Evans, Meirwyn C.; Dyson, Paul J.

In: Microbiology, Vol. 148, No. 2, 02.2002, p. 381-390.

Research output: Contribution to journalArticle

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N2 - Transcriptional activation of the thiostrepton-inducible promoter, ptipA, in Streptomyces lividans is mediated by TipAL. This transcriptional activator belongs to the MerR/SoxR family that characteristically binds an operator sequence located between the -10 and -35 hexamers normally occupied by RNA polymerase. As for the Escherichia coli merT promoter, the ptipA hexamers are separated by a long 19 bp spacer and hence a topological transition of the DNA is likely to be a requisite for alignment with RNA polymerase. Growth conditions that could facilitate this conformational change were investigated using transcriptional fusions of ptipA with reporter genes. Adjustment of growth medium osmolarity led to increased and prolonged TipAL-dependent expression, both with and without the inducer, thiostrepton. These effects correlated with increases in negative DNA supercoiling. Moreover, an inability to induce the promoter with thiostrepton in strain TK64 was corrected by increasing the concentration of osmolyte, compensating for an apparent reduced level of negative DNA supercoiling in the strain. Prolonging the time of activation of tipA in the wild-type by manipulating growth conditions revealed that mycelial autolysis could be induced by thiostrepton in 4-d-old cultures.

AB - Transcriptional activation of the thiostrepton-inducible promoter, ptipA, in Streptomyces lividans is mediated by TipAL. This transcriptional activator belongs to the MerR/SoxR family that characteristically binds an operator sequence located between the -10 and -35 hexamers normally occupied by RNA polymerase. As for the Escherichia coli merT promoter, the ptipA hexamers are separated by a long 19 bp spacer and hence a topological transition of the DNA is likely to be a requisite for alignment with RNA polymerase. Growth conditions that could facilitate this conformational change were investigated using transcriptional fusions of ptipA with reporter genes. Adjustment of growth medium osmolarity led to increased and prolonged TipAL-dependent expression, both with and without the inducer, thiostrepton. These effects correlated with increases in negative DNA supercoiling. Moreover, an inability to induce the promoter with thiostrepton in strain TK64 was corrected by increasing the concentration of osmolyte, compensating for an apparent reduced level of negative DNA supercoiling in the strain. Prolonging the time of activation of tipA in the wild-type by manipulating growth conditions revealed that mycelial autolysis could be induced by thiostrepton in 4-d-old cultures.

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