Increased Synthesis of an Escherichia coli Membrane Protein Suppresses F Exclusion of Bacteriophage T7

Wang, Wen Fang; Margolin, William; Molineux, Ian J.

doi:10.1006/jmbi.1999.3088

Cited by 5 publications

(1 citation statement)

References 47 publications

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“…The maximum measured Syn5 burst size of 30 virions per cell was recorded when cultures of WH8109 were in log phase and at a concentration of approximately 1×10 8 cells/ml (data not shown). This burst size is about a fourth of the reported burst sizes for phages like T7 (120 phage/cell) 23 and phi-YeO-3 (132 phage/cell) 24 .…”

Section: Conditions For Propagating Cyanophage At High Titermentioning

confidence: 59%

Genome Sequence, Structural Proteins, and Capsid Organization of the Cyanophage Syn5: A “Horned” Bacteriophage of Marine Synechococcus

Pope

Weigele

Chang

et al. 2007

Journal of Molecular Biology

115

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Marine Synechococcus spp and marine Prochlorococcus spp are numerically dominant photoautotrophs in the open oceans and contributors to the global carbon cycle. Syn5 is a short-tailed cyanophage isolated from the Sargasso Sea on Synechococcus strain WH8109. Syn5 has been grown in WH8109 to high titer in the laboratory and purified and concentrated retaining infectivity. Genome sequencing and annotation of Syn5 revealed that the linear genome is 46,214bp with a 237bp terminal direct repeat. Sixty-one open reading frames (ORFs) were identified. Based on genomic organization and sequence similarity to known protein sequences within GenBank, Syn5 shares features with T7-like phages. The presence of a putative integrase suggests access to a temperate life-cycle. Assignment of eleven ORFs to structural proteins found within the phage virion was confirmed by mass-spectrometry and N-terminal sequencing. Eight of these identified structural proteins exhibited amino acid sequence similarity to enteric phage proteins. The remaining three virion proteins did not resemble any known phage sequences in GenBank as of August 2006. Cryoelectron micrographs of purified Syn5 virions revealed that the capsid has a single "horn", a novel fibrous structure protruding from the opposing end of the capsid from the tail of the virion. The tail appendage displayed an apparent three-fold rather than six-fold symmetry. An 18Å-resolution icosahedral reconstruction of the capsid revealed a T=7 lattice, but with an unusual pattern of surface knobs. This phage/host system should allow detailed investigation of the physiology and biochemistry of phage propagation in marine photosynthetic bacteria.

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Section: Conditions For Propagating Cyanophage At High Titermentioning

confidence: 59%

Genome Sequence, Structural Proteins, and Capsid Organization of the Cyanophage Syn5: A “Horned” Bacteriophage of Marine Synechococcus

Pope

Weigele

Chang

et al. 2007

Journal of Molecular Biology

115

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Isolation and Identification of fxsA, an Escherichia coli Gene that can Suppress F Exclusion of Bacteriophage T7

Wang

Cheng

Molineux

1999

Journal of Molecular Biology

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An online monitoring system based on a synthetic sigma32-dependent tandem promoter for visualization of insoluble proteins in the cytoplasm of Escherichia coli

Kraft

Knüpfer

Wenderoth

et al. 2007

Appl Microbiol Biotechnol

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The expression of heterologous proteins in the cytoplasm of Escherichia coli is often accompanied by limitations resulting in uncontrollable fermentation processes, increased rates of cell lysis, and thus limited yields of target protein. To deal with these problems, reporter tools are required to improve the folding properties of recombinant protein. In this work, the well-known sigma(32)-dependent promoters ibpAB and fxsA were linked in a tandem promoter (ibpfxs), fused with the luciferase reporter gene lucA to allow enhanced monitoring of the formation of misfolded proteins and their aggregates in E. coli cells. Overexpression of MalE31, a folding-defective variant of the maltose-binding protein, and other partially insoluble heterologous proteins showed that the lucA reporter gene was activated in the presence of these misfolded proteins. Contrary to this, the absence of damaged proteins or overexpression of mostly soluble proteins led to a reduced level of luciferase induction. Through performing expression of aggregation-prone proteins, we were able to demonstrate that the ibpfxs::lucA reporter unit is 2.5-4.5 times stronger than the single reporter units ibp::lucA and fxs::lucA. Data of misfolding studies showed that this reporter system provides an adequate tool for in vivo folding studies in E. coli from microtiter up to fermentation scales.

show abstract

Increased Synthesis of an Escherichia coli Membrane Protein Suppresses F Exclusion of Bacteriophage T7

Cited by 5 publications

References 47 publications

Genome Sequence, Structural Proteins, and Capsid Organization of the Cyanophage Syn5: A “Horned” Bacteriophage of Marine Synechococcus

Genome Sequence, Structural Proteins, and Capsid Organization of the Cyanophage Syn5: A “Horned” Bacteriophage of Marine Synechococcus

Isolation and Identification of fxsA, an Escherichia coli Gene that can Suppress F Exclusion of Bacteriophage T7

An online monitoring system based on a synthetic sigma32-dependent tandem promoter for visualization of insoluble proteins in the cytoplasm of Escherichia coli

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