Fluorescence Bleaching Reveals Asymmetric Compartment Formation Prior to Cell Division in Caulobacter
Asymmetric cell division in Caulobacter crescentus yields daughter cells that have different cell fates. Compartmentalization of the predivisional cell is a critical event in the establishment of the differential distribution of regulatory factors that specify cell fate. To determine when during the...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 2003-07, Vol.100 (14), p.8235-8240 |
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| creator | Judd, Ellen M. Ryan, Kathleen R. Moerner, W. E. Shapiro, Lucy McAdams, Harley H. |
| description | Asymmetric cell division in Caulobacter crescentus yields daughter cells that have different cell fates. Compartmentalization of the predivisional cell is a critical event in the establishment of the differential distribution of regulatory factors that specify cell fate. To determine when during the cell cycle the cytoplasm is compartmentalized so that cytoplasmic proteins can no longer diffuse between the two nascent progeny cell compartments, we designed a fluorescence loss in photobleaching assay. Individual cells containing enhanced GFP were exposed to a bleaching laser pulse tightly focused at one cell pole. In compartmentalized cells, fluorescence disappears only in the compartment receiving the bleaching beam; in noncompartmentalized cells, fluorescence disappears from the entire cell. In a 135-min cell cycle, the cells were compartmentalized 18 ± 5 min before the progeny cells separated. Clearance of the 22000 CtrA master transcriptional regulator molecules from the stalked portion of the predivisional cell is a controlling element of Caulobacter asymmetry. Monitoring of a fluorescent marker for CtrA showed that the differential degradation of CtrA in the nascent stalk cell compartment occurs only after the cytoplasm is compartmentalized. |
| doi_str_mv | 10.1073/pnas.1433105100 |
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E. ; Shapiro, Lucy ; McAdams, Harley H.</creator><creatorcontrib>Judd, Ellen M. ; Ryan, Kathleen R. ; Moerner, W. E. ; Shapiro, Lucy ; McAdams, Harley H.</creatorcontrib><description>Asymmetric cell division in Caulobacter crescentus yields daughter cells that have different cell fates. Compartmentalization of the predivisional cell is a critical event in the establishment of the differential distribution of regulatory factors that specify cell fate. To determine when during the cell cycle the cytoplasm is compartmentalized so that cytoplasmic proteins can no longer diffuse between the two nascent progeny cell compartments, we designed a fluorescence loss in photobleaching assay. Individual cells containing enhanced GFP were exposed to a bleaching laser pulse tightly focused at one cell pole. In compartmentalized cells, fluorescence disappears only in the compartment receiving the bleaching beam; in noncompartmentalized cells, fluorescence disappears from the entire cell. In a 135-min cell cycle, the cells were compartmentalized 18 ± 5 min before the progeny cells separated. Clearance of the 22000 CtrA master transcriptional regulator molecules from the stalked portion of the predivisional cell is a controlling element of Caulobacter asymmetry. Monitoring of a fluorescent marker for CtrA showed that the differential degradation of CtrA in the nascent stalk cell compartment occurs only after the cytoplasm is compartmentalized.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.1433105100</identifier><identifier>PMID: 12824468</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>Bacterial Proteins - biosynthesis ; Bacterial Proteins - genetics ; Bacterial Proteins - physiology ; Biological Sciences ; Bleaching ; Caulobacter ; Caulobacter crescentus - cytology ; Caulobacter crescentus - genetics ; Caulobacter crescentus - radiation effects ; Cell Compartmentation ; Cell Cycle ; Cell Division ; Cell separation ; Computer Simulation ; Diffusion ; DNA Replication ; DNA-Binding Proteins - physiology ; Fluorescence ; Gene Expression Regulation, Bacterial ; Green Fluorescent Proteins ; Laser beams ; Lasers ; Luminescent Proteins - radiation effects ; Models, Biological ; Molecules ; Mother cells ; Photochemistry ; Transcription Factors - physiology ; Transcription, Genetic</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2003-07, Vol.100 (14), p.8235-8240</ispartof><rights>Copyright 1993-2003 National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences Jul 8, 2003</rights><rights>Copyright © 2003, The National Academy of Sciences 2003</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c524t-cdef161dfafa952246eecc354f66e727077d2db1bda8375143531737bd5645a53</citedby><cites>FETCH-LOGICAL-c524t-cdef161dfafa952246eecc354f66e727077d2db1bda8375143531737bd5645a53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/100/14.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/3139910$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/3139910$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,723,776,780,799,881,27903,27904,53770,53772,57996,58229</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12824468$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Judd, Ellen M.</creatorcontrib><creatorcontrib>Ryan, Kathleen R.</creatorcontrib><creatorcontrib>Moerner, W. E.</creatorcontrib><creatorcontrib>Shapiro, Lucy</creatorcontrib><creatorcontrib>McAdams, Harley H.</creatorcontrib><title>Fluorescence Bleaching Reveals Asymmetric Compartment Formation Prior to Cell Division in Caulobacter</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Asymmetric cell division in Caulobacter crescentus yields daughter cells that have different cell fates. Compartmentalization of the predivisional cell is a critical event in the establishment of the differential distribution of regulatory factors that specify cell fate. To determine when during the cell cycle the cytoplasm is compartmentalized so that cytoplasmic proteins can no longer diffuse between the two nascent progeny cell compartments, we designed a fluorescence loss in photobleaching assay. Individual cells containing enhanced GFP were exposed to a bleaching laser pulse tightly focused at one cell pole. In compartmentalized cells, fluorescence disappears only in the compartment receiving the bleaching beam; in noncompartmentalized cells, fluorescence disappears from the entire cell. In a 135-min cell cycle, the cells were compartmentalized 18 ± 5 min before the progeny cells separated. Clearance of the 22000 CtrA master transcriptional regulator molecules from the stalked portion of the predivisional cell is a controlling element of Caulobacter asymmetry. Monitoring of a fluorescent marker for CtrA showed that the differential degradation of CtrA in the nascent stalk cell compartment occurs only after the cytoplasm is compartmentalized.</description><subject>Bacterial Proteins - biosynthesis</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - physiology</subject><subject>Biological Sciences</subject><subject>Bleaching</subject><subject>Caulobacter</subject><subject>Caulobacter crescentus - cytology</subject><subject>Caulobacter crescentus - genetics</subject><subject>Caulobacter crescentus - radiation effects</subject><subject>Cell Compartmentation</subject><subject>Cell Cycle</subject><subject>Cell Division</subject><subject>Cell separation</subject><subject>Computer Simulation</subject><subject>Diffusion</subject><subject>DNA Replication</subject><subject>DNA-Binding Proteins - physiology</subject><subject>Fluorescence</subject><subject>Gene Expression Regulation, Bacterial</subject><subject>Green Fluorescent Proteins</subject><subject>Laser beams</subject><subject>Lasers</subject><subject>Luminescent Proteins - radiation effects</subject><subject>Models, Biological</subject><subject>Molecules</subject><subject>Mother cells</subject><subject>Photochemistry</subject><subject>Transcription Factors - physiology</subject><subject>Transcription, Genetic</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1v1DAQxSMEotvCmQsCqwfEJa2_nRw4lMACUiUQgrPlJJPWKydebGdF_3sc7aoLHOBkafx7T_PmFcUzgi8IVuxyO5l4QThjBAuC8YNiRXBNSslr_LBYYUxVWXHKT4rTGDcY41pU-HFxQmhFOZfVqoC1m32A2MHUAXrrwHS3drpBX2EHxkV0Fe_GEVKwHWr8uDUhjTAltPZhNMn6CX0J1geUPGrAOfTO7mxcxnZCjZmdb02XIDwpHg3ZDZ4e3rPi-_r9t-Zjef35w6fm6rrsBOWp7HoYiCT9YAZTC0q5BOg6JvggJSiqsFI97VvS9qZiSuTcghHFVNsLyYUR7Kx4s_fdzu0IfQ6VgnF6G-xowp32xuo_fyZ7q2_8ThMpKaFZ_-qgD_7HDDHp0ebTOGcm8HPUinEhWKX-C5JK1UzKBTz_C9z4OUz5CJpiwmpCaZWhyz3UBR9jgOF-Y4L10rNeetbHnrPixe9Bj_yh2Ay8PACL8miX_biuKFtu9frfhB5m5xL8TBl9vkc3Mflwz7K8f00w-wUzksbB</recordid><startdate>20030708</startdate><enddate>20030708</enddate><creator>Judd, Ellen M.</creator><creator>Ryan, Kathleen R.</creator><creator>Moerner, W. 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E.</au><au>Shapiro, Lucy</au><au>McAdams, Harley H.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Fluorescence Bleaching Reveals Asymmetric Compartment Formation Prior to Cell Division in Caulobacter</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>2003-07-08</date><risdate>2003</risdate><volume>100</volume><issue>14</issue><spage>8235</spage><epage>8240</epage><pages>8235-8240</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>Asymmetric cell division in Caulobacter crescentus yields daughter cells that have different cell fates. Compartmentalization of the predivisional cell is a critical event in the establishment of the differential distribution of regulatory factors that specify cell fate. To determine when during the cell cycle the cytoplasm is compartmentalized so that cytoplasmic proteins can no longer diffuse between the two nascent progeny cell compartments, we designed a fluorescence loss in photobleaching assay. Individual cells containing enhanced GFP were exposed to a bleaching laser pulse tightly focused at one cell pole. In compartmentalized cells, fluorescence disappears only in the compartment receiving the bleaching beam; in noncompartmentalized cells, fluorescence disappears from the entire cell. In a 135-min cell cycle, the cells were compartmentalized 18 ± 5 min before the progeny cells separated. Clearance of the 22000 CtrA master transcriptional regulator molecules from the stalked portion of the predivisional cell is a controlling element of Caulobacter asymmetry. 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| subjects | Bacterial Proteins - biosynthesis Bacterial Proteins - genetics Bacterial Proteins - physiology Biological Sciences Bleaching Caulobacter Caulobacter crescentus - cytology Caulobacter crescentus - genetics Caulobacter crescentus - radiation effects Cell Compartmentation Cell Cycle Cell Division Cell separation Computer Simulation Diffusion DNA Replication DNA-Binding Proteins - physiology Fluorescence Gene Expression Regulation, Bacterial Green Fluorescent Proteins Laser beams Lasers Luminescent Proteins - radiation effects Models, Biological Molecules Mother cells Photochemistry Transcription Factors - physiology Transcription, Genetic |
| title | Fluorescence Bleaching Reveals Asymmetric Compartment Formation Prior to Cell Division in Caulobacter |
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