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On the spatial organization of mRNA, plasmids, and ribosomes in a bacterial host overexpressing membrane proteins

Journal Article


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Abstract


  • By using fluorescence imaging, we provide a time-resolved single-cell view on coupled defects in transcription, translation, and growth during expression of heterologous membrane proteins in Lactococcus lactis. Transcripts encoding poorly produced membrane proteins accumulate in mRNA-dense bodies at the cell poles, whereas transcripts of a well-expressed homologous membrane protein show membrane-proximal localization in a translation-dependent fashion. The presence of the aberrant polar mRNA foci correlates with cessation of cell division, which is restored once these bodies are cleared. In addition, activation of the heat-shock response and a loss of nucleoid-occluded ribosomes are observed. We show that the presence of a native-like N-terminal domain is key to SRP-dependent membrane localization and successful production of membrane proteins. The work presented gives new insights and detailed understanding of aberrant membrane protein biogenesis, which can be used for strategies to optimize membrane protein production.

Publication Date


  • 2016

Citation


  • van Gijtenbeek, L. A., Robinson, A., van Oijen, A. M., Poolman, B. & Kok, J. (2016). On the spatial organization of mRNA, plasmids, and ribosomes in a bacterial host overexpressing membrane proteins. PLoS Genetics, 12 (12), e1006523-1-e1006523-27.

Scopus Eid


  • 2-s2.0-85007560028

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=5530&context=smhpapers

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/4506

Has Global Citation Frequency


Start Page


  • e1006523-1

End Page


  • e1006523-27

Volume


  • 12

Issue


  • 12

Place Of Publication


  • United States

Abstract


  • By using fluorescence imaging, we provide a time-resolved single-cell view on coupled defects in transcription, translation, and growth during expression of heterologous membrane proteins in Lactococcus lactis. Transcripts encoding poorly produced membrane proteins accumulate in mRNA-dense bodies at the cell poles, whereas transcripts of a well-expressed homologous membrane protein show membrane-proximal localization in a translation-dependent fashion. The presence of the aberrant polar mRNA foci correlates with cessation of cell division, which is restored once these bodies are cleared. In addition, activation of the heat-shock response and a loss of nucleoid-occluded ribosomes are observed. We show that the presence of a native-like N-terminal domain is key to SRP-dependent membrane localization and successful production of membrane proteins. The work presented gives new insights and detailed understanding of aberrant membrane protein biogenesis, which can be used for strategies to optimize membrane protein production.

Publication Date


  • 2016

Citation


  • van Gijtenbeek, L. A., Robinson, A., van Oijen, A. M., Poolman, B. & Kok, J. (2016). On the spatial organization of mRNA, plasmids, and ribosomes in a bacterial host overexpressing membrane proteins. PLoS Genetics, 12 (12), e1006523-1-e1006523-27.

Scopus Eid


  • 2-s2.0-85007560028

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=5530&context=smhpapers

Ro Metadata Url


  • http://ro.uow.edu.au/smhpapers/4506

Has Global Citation Frequency


Start Page


  • e1006523-1

End Page


  • e1006523-27

Volume


  • 12

Issue


  • 12

Place Of Publication


  • United States