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Full-Text Articles in Social and Behavioral Sciences
On The Spatial Organization Of Mrna, Plasmids, And Ribosomes In A Bacterial Host Overexpressing Membrane Proteins, Lieke A. Van Gijtenbeek, Andrew Robinson, Antoine M. Van Oijen, Bert Poolman, Jan Kok
On The Spatial Organization Of Mrna, Plasmids, And Ribosomes In A Bacterial Host Overexpressing Membrane Proteins, Lieke A. Van Gijtenbeek, Andrew Robinson, Antoine M. Van Oijen, Bert Poolman, Jan Kok
Faculty of Science, Medicine and Health - Papers: part A
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 …
Single-Molecule Specific Mislocalization Of Red Fluorescent Proteins In Live Escherichia Coli, Harshad Ghodke, Victor E. A Caldas, Christiaan M. Punter, Antoine M. Van Oijen, Andrew Robinson
Single-Molecule Specific Mislocalization Of Red Fluorescent Proteins In Live Escherichia Coli, Harshad Ghodke, Victor E. A Caldas, Christiaan M. Punter, Antoine M. Van Oijen, Andrew Robinson
Faculty of Science, Medicine and Health - Papers: part A
Tagging of individual proteins with genetically encoded fluorescent proteins (FPs) has been used extensively to study localization and interactions in live cells. Recent developments in single-molecule localization microscopy have enabled the dynamic visualization of individual tagged proteins inside living cells. However, tagging proteins with FPs is not without problems: formation of insoluble aggregates and inhibition of native functions of the protein are well-known issues. Previously reported artifacts manifest themselves at all expression levels of the FP-tagged proteins, making the design of control experiments relatively straightforward. Here, we describe a previously uncharacterized mislocalization artifact of Entacmaea quadricolor red fluorescent protein variants …