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Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate
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| Zeitschriftentitel: | Proceedings of the National Academy of Sciences |
|---|---|
| Personen und Körperschaften: | , |
| In: | Proceedings of the National Academy of Sciences, 76, 1979, 4, S. 1726-1730 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Proceedings of the National Academy of Sciences
|
| Schlagwörter: |
| author_facet |
Primakoff, Paul Artz, Stanley W. Primakoff, Paul Artz, Stanley W. |
|---|---|
| author |
Primakoff, Paul Artz, Stanley W. |
| spellingShingle |
Primakoff, Paul Artz, Stanley W. Proceedings of the National Academy of Sciences Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate Multidisciplinary |
| author_sort |
primakoff, paul |
| spelling |
Primakoff, Paul Artz, Stanley W. 0027-8424 1091-6490 Proceedings of the National Academy of Sciences Multidisciplinary http://dx.doi.org/10.1073/pnas.76.4.1726 <jats:p> Maximal expression of the <jats:italic>Escherichia coli</jats:italic> lactose operon in a coupled <jats:italic>in vitro</jats:italic> transcription-translation system from a <jats:italic>Salmonella typhimurium relA</jats:italic> mutant was strongly dependent upon addition of guanosine 5′-diphosphate 3′-diphosphate (ppGpp). Without added ppGpp, at saturating 3′,5′-cyclic AMP (cAMP) concentrations, synthesis of β-galactosidase (β-D-galactoside galactohydrolase, EC 3.2.1.23) was reproducibly only 5-7% of that which can be obtained with 0.5-0.8 mM ppGpp. Experiments in which transcription was uncoupled from translation indicated that this 14- to 20-fold stimulation by ppGpp occurred at the level of transcription. When coupled β-galactosidase synthesis was primed with a template containing a well-characterized mutant <jats:italic>lac</jats:italic> promoter ( <jats:italic>lacP</jats:italic> <jats:sup>r</jats:sup> L8UV5), the dependence on ppGpp was greatly reduced. This result provides an important experimental control previously unavailable for verifying the significance of ppGpp effects on gene regulation <jats:italic>in vitro</jats:italic> ; it indicates that activation of <jats:italic>lacP</jats:italic> <jats:sup>+</jats:sup> expression by ppGpp is specifically an effect of increased transcription initiations. Furthermore, the large ppGpp stimulation of <jats:italic>lacP</jats:italic> <jats:sup>+</jats:sup> DNA enabled the level of expression of this template to approach that of <jats:italic>lacP</jats:italic> <jats:sup>r</jats:sup> L8UV5 DNA, an observation expected from results <jats:italic>in vivo</jats:italic> but not obtained with other transcription-translation systems <jats:italic>in vitro</jats:italic> . The importance of these results is considered with respect to previous ideas on the physiological role of ppGpp as a supercontrol molecule in bacterial regulation. </jats:p> Positive control of <i>lac</i> operon expression in <i>vitro</i> by guanosine 5′-diphosphate 3′-diphosphate Proceedings of the National Academy of Sciences |
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10.1073/pnas.76.4.1726 |
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Proceedings of the National Academy of Sciences, 1979 |
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Proceedings of the National Academy of Sciences, 1979 |
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1091-6490 0027-8424 |
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primakoff1979positivecontroloflacoperonexpressioninvitrobyguanosine5diphosphate3diphosphate |
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1979 |
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Proceedings of the National Academy of Sciences |
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ai |
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Proceedings of the National Academy of Sciences |
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49 |
| title |
Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| title_unstemmed |
Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| title_full |
Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| title_fullStr |
Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| title_full_unstemmed |
Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| title_short |
Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| title_sort |
positive control of
<i>lac</i>
operon expression in
<i>vitro</i>
by guanosine 5′-diphosphate 3′-diphosphate |
| topic |
Multidisciplinary |
| url |
http://dx.doi.org/10.1073/pnas.76.4.1726 |
| publishDate |
1979 |
| physical |
1726-1730 |
| description |
<jats:p>
Maximal expression of the
<jats:italic>Escherichia coli</jats:italic>
lactose operon in a coupled
<jats:italic>in vitro</jats:italic>
transcription-translation system from a
<jats:italic>Salmonella typhimurium relA</jats:italic>
mutant was strongly dependent upon addition of guanosine 5′-diphosphate 3′-diphosphate (ppGpp). Without added ppGpp, at saturating 3′,5′-cyclic AMP (cAMP) concentrations, synthesis of β-galactosidase (β-D-galactoside galactohydrolase, EC 3.2.1.23) was reproducibly only 5-7% of that which can be obtained with 0.5-0.8 mM ppGpp. Experiments in which transcription was uncoupled from translation indicated that this 14- to 20-fold stimulation by ppGpp occurred at the level of transcription. When coupled β-galactosidase synthesis was primed with a template containing a well-characterized mutant
<jats:italic>lac</jats:italic>
promoter (
<jats:italic>lacP</jats:italic>
<jats:sup>r</jats:sup>
L8UV5), the dependence on ppGpp was greatly reduced. This result provides an important experimental control previously unavailable for verifying the significance of ppGpp effects on gene regulation
<jats:italic>in vitro</jats:italic>
; it indicates that activation of
<jats:italic>lacP</jats:italic>
<jats:sup>+</jats:sup>
expression by ppGpp is specifically an effect of increased transcription initiations. Furthermore, the large ppGpp stimulation of
<jats:italic>lacP</jats:italic>
<jats:sup>+</jats:sup>
DNA enabled the level of expression of this template to approach that of
<jats:italic>lacP</jats:italic>
<jats:sup>r</jats:sup>
L8UV5 DNA, an observation expected from results
<jats:italic>in vivo</jats:italic>
but not obtained with other transcription-translation systems
<jats:italic>in vitro</jats:italic>
. The importance of these results is considered with respect to previous ideas on the physiological role of ppGpp as a supercontrol molecule in bacterial regulation.
</jats:p> |
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| author | Primakoff, Paul, Artz, Stanley W. |
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| description | <jats:p> Maximal expression of the <jats:italic>Escherichia coli</jats:italic> lactose operon in a coupled <jats:italic>in vitro</jats:italic> transcription-translation system from a <jats:italic>Salmonella typhimurium relA</jats:italic> mutant was strongly dependent upon addition of guanosine 5′-diphosphate 3′-diphosphate (ppGpp). Without added ppGpp, at saturating 3′,5′-cyclic AMP (cAMP) concentrations, synthesis of β-galactosidase (β-D-galactoside galactohydrolase, EC 3.2.1.23) was reproducibly only 5-7% of that which can be obtained with 0.5-0.8 mM ppGpp. Experiments in which transcription was uncoupled from translation indicated that this 14- to 20-fold stimulation by ppGpp occurred at the level of transcription. When coupled β-galactosidase synthesis was primed with a template containing a well-characterized mutant <jats:italic>lac</jats:italic> promoter ( <jats:italic>lacP</jats:italic> <jats:sup>r</jats:sup> L8UV5), the dependence on ppGpp was greatly reduced. This result provides an important experimental control previously unavailable for verifying the significance of ppGpp effects on gene regulation <jats:italic>in vitro</jats:italic> ; it indicates that activation of <jats:italic>lacP</jats:italic> <jats:sup>+</jats:sup> expression by ppGpp is specifically an effect of increased transcription initiations. Furthermore, the large ppGpp stimulation of <jats:italic>lacP</jats:italic> <jats:sup>+</jats:sup> DNA enabled the level of expression of this template to approach that of <jats:italic>lacP</jats:italic> <jats:sup>r</jats:sup> L8UV5 DNA, an observation expected from results <jats:italic>in vivo</jats:italic> but not obtained with other transcription-translation systems <jats:italic>in vitro</jats:italic> . The importance of these results is considered with respect to previous ideas on the physiological role of ppGpp as a supercontrol molecule in bacterial regulation. </jats:p> |
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| imprint | Proceedings of the National Academy of Sciences, 1979 |
| imprint_str_mv | Proceedings of the National Academy of Sciences, 1979 |
| institution | DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1, DE-L229, DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161 |
| issn | 1091-6490, 0027-8424 |
| issn_str_mv | 1091-6490, 0027-8424 |
| language | English |
| last_indexed | 2024-03-01T15:06:53.752Z |
| match_str | primakoff1979positivecontroloflacoperonexpressioninvitrobyguanosine5diphosphate3diphosphate |
| mega_collection | Proceedings of the National Academy of Sciences (CrossRef) |
| physical | 1726-1730 |
| publishDate | 1979 |
| publishDateSort | 1979 |
| publisher | Proceedings of the National Academy of Sciences |
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| recordtype | ai |
| series | Proceedings of the National Academy of Sciences |
| source_id | 49 |
| spelling | Primakoff, Paul Artz, Stanley W. 0027-8424 1091-6490 Proceedings of the National Academy of Sciences Multidisciplinary http://dx.doi.org/10.1073/pnas.76.4.1726 <jats:p> Maximal expression of the <jats:italic>Escherichia coli</jats:italic> lactose operon in a coupled <jats:italic>in vitro</jats:italic> transcription-translation system from a <jats:italic>Salmonella typhimurium relA</jats:italic> mutant was strongly dependent upon addition of guanosine 5′-diphosphate 3′-diphosphate (ppGpp). Without added ppGpp, at saturating 3′,5′-cyclic AMP (cAMP) concentrations, synthesis of β-galactosidase (β-D-galactoside galactohydrolase, EC 3.2.1.23) was reproducibly only 5-7% of that which can be obtained with 0.5-0.8 mM ppGpp. Experiments in which transcription was uncoupled from translation indicated that this 14- to 20-fold stimulation by ppGpp occurred at the level of transcription. When coupled β-galactosidase synthesis was primed with a template containing a well-characterized mutant <jats:italic>lac</jats:italic> promoter ( <jats:italic>lacP</jats:italic> <jats:sup>r</jats:sup> L8UV5), the dependence on ppGpp was greatly reduced. This result provides an important experimental control previously unavailable for verifying the significance of ppGpp effects on gene regulation <jats:italic>in vitro</jats:italic> ; it indicates that activation of <jats:italic>lacP</jats:italic> <jats:sup>+</jats:sup> expression by ppGpp is specifically an effect of increased transcription initiations. Furthermore, the large ppGpp stimulation of <jats:italic>lacP</jats:italic> <jats:sup>+</jats:sup> DNA enabled the level of expression of this template to approach that of <jats:italic>lacP</jats:italic> <jats:sup>r</jats:sup> L8UV5 DNA, an observation expected from results <jats:italic>in vivo</jats:italic> but not obtained with other transcription-translation systems <jats:italic>in vitro</jats:italic> . The importance of these results is considered with respect to previous ideas on the physiological role of ppGpp as a supercontrol molecule in bacterial regulation. </jats:p> Positive control of <i>lac</i> operon expression in <i>vitro</i> by guanosine 5′-diphosphate 3′-diphosphate Proceedings of the National Academy of Sciences |
| spellingShingle | Primakoff, Paul, Artz, Stanley W., Proceedings of the National Academy of Sciences, Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate, Multidisciplinary |
| title | Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| title_full | Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| title_fullStr | Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| title_full_unstemmed | Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| title_short | Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| title_sort | positive control of <i>lac</i> operon expression in <i>vitro</i> by guanosine 5′-diphosphate 3′-diphosphate |
| title_unstemmed | Positive control of lac operon expression in vitro by guanosine 5′-diphosphate 3′-diphosphate |
| topic | Multidisciplinary |
| url | http://dx.doi.org/10.1073/pnas.76.4.1726 |