Segregation of molecules at cell division reveals native protein localization
We introduce a nonintrusive method exploiting single-cell variability after cell division to validate protein localization. We found that Clp proteases, widely reported to form biologically relevant foci, were uniformly distributed in Escherichia coli cells, and that many commonly used fluorescent p...
| Main Authors: | , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2014
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| Online Access: | http://hdl.handle.net/1721.1/84680 |
| _version_ | 1826196947823755264 |
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| author | Landgraf, Dirk Okumus, Burak Chien, Peter Baker, Tania Paulsson, Johan |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Landgraf, Dirk Okumus, Burak Chien, Peter Baker, Tania Paulsson, Johan |
| author_sort | Landgraf, Dirk |
| collection | MIT |
| description | We introduce a nonintrusive method exploiting single-cell variability after cell division to validate protein localization. We found that Clp proteases, widely reported to form biologically relevant foci, were uniformly distributed in Escherichia coli cells, and that many commonly used fluorescent proteins caused severe mislocalization when fused to homo-oligomers. Retagging five other reportedly foci-forming proteins with the most monomeric fluorescent protein tested suggests that the foci were caused by the fluorescent tags. |
| first_indexed | 2024-09-23T10:40:25Z |
| format | Article |
| id | mit-1721.1/84680 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:40:25Z |
| publishDate | 2014 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/846802021-09-10T15:13:23Z Segregation of molecules at cell division reveals native protein localization Landgraf, Dirk Okumus, Burak Chien, Peter Baker, Tania Paulsson, Johan Massachusetts Institute of Technology. Department of Biology Whitehead Institute for Biomedical Research Baker, Tania Chien, Peter We introduce a nonintrusive method exploiting single-cell variability after cell division to validate protein localization. We found that Clp proteases, widely reported to form biologically relevant foci, were uniformly distributed in Escherichia coli cells, and that many commonly used fluorescent proteins caused severe mislocalization when fused to homo-oligomers. Retagging five other reportedly foci-forming proteins with the most monomeric fluorescent protein tested suggests that the foci were caused by the fluorescent tags. National Institutes of Health (U.S.) (grant GM081563) National Institutes of Health (U.S.) (grant GM095784) 2014-02-07T16:54:47Z 2014-02-07T16:54:47Z 2012-04 Article http://purl.org/eprint/type/JournalArticle 1548-7091 1548-7105 http://hdl.handle.net/1721.1/84680 Landgraf, Dirk, Burak Okumus, Peter Chien, Tania A Baker, and Johan Paulsson. “Segregation of molecules at cell division reveals native protein localization.” Nature Methods 9, no. 5 (April 8, 2012): 480-482. en_US http://dx.doi.org/10.1038/nmeth.1955 Nature Methods Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Nature Publishing Group PMC |
| spellingShingle | Landgraf, Dirk Okumus, Burak Chien, Peter Baker, Tania Paulsson, Johan Segregation of molecules at cell division reveals native protein localization |
| title | Segregation of molecules at cell division reveals native protein localization |
| title_full | Segregation of molecules at cell division reveals native protein localization |
| title_fullStr | Segregation of molecules at cell division reveals native protein localization |
| title_full_unstemmed | Segregation of molecules at cell division reveals native protein localization |
| title_short | Segregation of molecules at cell division reveals native protein localization |
| title_sort | segregation of molecules at cell division reveals native protein localization |
| url | http://hdl.handle.net/1721.1/84680 |
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