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ParB spreading on DNA requires cytidine triphosphate in vitro
In all living organisms, it is essential to transmit genetic information faithfully to the next generation. The SMC-ParAB- system is widely employed for chromosome segregation in bacteria. A DNA-binding protein ParB nucleates on sites and must associate with neighboring DNA, a process known as sprea...
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| Published in: | eLife 2020-02, Vol.9 |
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| creator | Jalal, Adam Sb Tran, Ngat T Le, Tung Bk |
| description | In all living organisms, it is essential to transmit genetic information faithfully to the next generation. The SMC-ParAB-
system is widely employed for chromosome segregation in bacteria. A DNA-binding protein ParB nucleates on
sites and must associate with neighboring DNA, a process known as spreading, to enable efficient chromosome segregation. Despite its importance, how the initial few ParB molecules nucleating at
sites recruit hundreds of further ParB to spread is not fully understood. Here, we reconstitute a
-dependent ParB spreading event using purified proteins from
and show that CTP is required for spreading. We further show that ParB spreading requires a closed DNA substrate, and a DNA-binding transcriptional regulator can act as a roadblock to attenuate spreading unidirectionally in vitro. Our biochemical reconstitutions recapitulate many observed in vivo properties of ParB and opens up avenues to investigate the interactions between ParB-
with ParA and SMC. |
| doi_str_mv | 10.7554/elife.53515 |
| format | article |
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system is widely employed for chromosome segregation in bacteria. A DNA-binding protein ParB nucleates on
sites and must associate with neighboring DNA, a process known as spreading, to enable efficient chromosome segregation. Despite its importance, how the initial few ParB molecules nucleating at
sites recruit hundreds of further ParB to spread is not fully understood. Here, we reconstitute a
-dependent ParB spreading event using purified proteins from
and show that CTP is required for spreading. We further show that ParB spreading requires a closed DNA substrate, and a DNA-binding transcriptional regulator can act as a roadblock to attenuate spreading unidirectionally in vitro. Our biochemical reconstitutions recapitulate many observed in vivo properties of ParB and opens up avenues to investigate the interactions between ParB-
with ParA and SMC.</description><identifier>ISSN: 2050-084X</identifier><identifier>EISSN: 2050-084X</identifier><identifier>DOI: 10.7554/elife.53515</identifier><identifier>PMID: 32077854</identifier><language>eng</language><publisher>England: eLife Science Publications, Ltd</publisher><subject>Bacteria ; Bacterial genetics ; Bacterial Proteins - metabolism ; Binding proteins ; Binding sites ; Caulobacter crescentus ; Caulobacter crescentus - metabolism ; chromosome organization ; chromosome segregation ; Chromosomes ; Chromosomes and Gene Expression ; CTP ; Cytidine triphosphate ; Cytidine Triphosphate - metabolism ; Deoxyribonucleic acid ; DNA ; DNA Primase - metabolism ; DNA, Bacterial - metabolism ; DNA-binding protein ; DNA-Binding Proteins - metabolism ; Hydrolysis ; Microbiology and Infectious Disease ; ParB-parS ; Protein binding ; Proteins ; Spreading ; Transcription ; Transcription (Genetics)</subject><ispartof>eLife, 2020-02, Vol.9</ispartof><rights>2020, Jalal et al.</rights><rights>COPYRIGHT 2020 eLife Science Publications, Ltd.</rights><rights>2020, Jalal et al. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2020, Jalal et al 2020 Jalal et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c642t-1da2c81e5fbd47e4d48830a3d5ce4da6a344bd7c67dd4ce591bb23fd8519c6e13</citedby><cites>FETCH-LOGICAL-c642t-1da2c81e5fbd47e4d48830a3d5ce4da6a344bd7c67dd4ce591bb23fd8519c6e13</cites><orcidid>0000-0001-7794-8834 ; 0000-0002-7186-3976 ; 0000-0003-4764-8851</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/2384740535/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/2384740535?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,25753,27924,27925,37012,37013,44590,53791,53793,75126</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32077854$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jalal, Adam Sb</creatorcontrib><creatorcontrib>Tran, Ngat T</creatorcontrib><creatorcontrib>Le, Tung Bk</creatorcontrib><title>ParB spreading on DNA requires cytidine triphosphate in vitro</title><title>eLife</title><addtitle>Elife</addtitle><description>In all living organisms, it is essential to transmit genetic information faithfully to the next generation. The SMC-ParAB-
system is widely employed for chromosome segregation in bacteria. A DNA-binding protein ParB nucleates on
sites and must associate with neighboring DNA, a process known as spreading, to enable efficient chromosome segregation. Despite its importance, how the initial few ParB molecules nucleating at
sites recruit hundreds of further ParB to spread is not fully understood. Here, we reconstitute a
-dependent ParB spreading event using purified proteins from
and show that CTP is required for spreading. We further show that ParB spreading requires a closed DNA substrate, and a DNA-binding transcriptional regulator can act as a roadblock to attenuate spreading unidirectionally in vitro. Our biochemical reconstitutions recapitulate many observed in vivo properties of ParB and opens up avenues to investigate the interactions between ParB-
with ParA and SMC.</description><subject>Bacteria</subject><subject>Bacterial genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Binding proteins</subject><subject>Binding sites</subject><subject>Caulobacter crescentus</subject><subject>Caulobacter crescentus - metabolism</subject><subject>chromosome organization</subject><subject>chromosome segregation</subject><subject>Chromosomes</subject><subject>Chromosomes and Gene Expression</subject><subject>CTP</subject><subject>Cytidine triphosphate</subject><subject>Cytidine Triphosphate - metabolism</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA Primase - metabolism</subject><subject>DNA, Bacterial - metabolism</subject><subject>DNA-binding protein</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Hydrolysis</subject><subject>Microbiology and Infectious Disease</subject><subject>ParB-parS</subject><subject>Protein binding</subject><subject>Proteins</subject><subject>Spreading</subject><subject>Transcription</subject><subject>Transcription (Genetics)</subject><issn>2050-084X</issn><issn>2050-084X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNptkltvFCEUgCfGxjZtn3w3k_iiMbvCAAM8aLLW2yYbNV4S3wgDZ3bZzA5TYBr772W7tXaM8MDt4wMOpygeYzTnjNGX0LkW5owwzB4UJxViaIYE_fnwXv-4OI9xi3LhVAgsHxXHpEKcC0ZPildfdHhTxiGAtq5fl74v335alAEuRxcgluY6ubwAZQpu2Pg4bHSC0vXllUvBnxVHre4inN-2p8WP9---X3ycrT5_WF4sVjNT0yrNsNWVERhY21jKgdp8D4I0sczkga41obSx3NTcWmqASdw0FWmtYFiaGjA5LZYHr_V6q4bgdjpcK6-dupnwYa10SM50oIQEUzOJJG4lzRpZUyxEK4BYWUEls-v1wTWMzQ6sgT4F3U2k05XebdTaXymOGJFyL3h2Kwj-coSY1M5FA12ne_BjVBVhkkhG-P7eT_9Bt34MfQ5VpgTlNCvZX2qt8wNc3_p8rtlL1aLGNUb5p6pMzf9D5Wph54zvoXV5frLh-WRDZhL8Sms9xqiW375O2RcH1gQfY4D2Lh4YqX2iKVjlRFM3iZbpJ_dDeMf-SSvyG8AKy8M</recordid><startdate>20200220</startdate><enddate>20200220</enddate><creator>Jalal, Adam Sb</creator><creator>Tran, Ngat T</creator><creator>Le, Tung Bk</creator><general>eLife Science Publications, Ltd</general><general>eLife Sciences Publications Ltd</general><general>eLife Sciences Publications, Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>ISR</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0001-7794-8834</orcidid><orcidid>https://orcid.org/0000-0002-7186-3976</orcidid><orcidid>https://orcid.org/0000-0003-4764-8851</orcidid></search><sort><creationdate>20200220</creationdate><title>ParB spreading on DNA requires cytidine triphosphate in vitro</title><author>Jalal, Adam Sb ; 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The SMC-ParAB-
system is widely employed for chromosome segregation in bacteria. A DNA-binding protein ParB nucleates on
sites and must associate with neighboring DNA, a process known as spreading, to enable efficient chromosome segregation. Despite its importance, how the initial few ParB molecules nucleating at
sites recruit hundreds of further ParB to spread is not fully understood. Here, we reconstitute a
-dependent ParB spreading event using purified proteins from
and show that CTP is required for spreading. We further show that ParB spreading requires a closed DNA substrate, and a DNA-binding transcriptional regulator can act as a roadblock to attenuate spreading unidirectionally in vitro. Our biochemical reconstitutions recapitulate many observed in vivo properties of ParB and opens up avenues to investigate the interactions between ParB-
with ParA and SMC.</abstract><cop>England</cop><pub>eLife Science Publications, Ltd</pub><pmid>32077854</pmid><doi>10.7554/elife.53515</doi><orcidid>https://orcid.org/0000-0001-7794-8834</orcidid><orcidid>https://orcid.org/0000-0002-7186-3976</orcidid><orcidid>https://orcid.org/0000-0003-4764-8851</orcidid><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 2050-084X |
| ispartof | eLife, 2020-02, Vol.9 |
| issn | 2050-084X 2050-084X |
| language | eng |
| recordid | cdi_doaj_primary_oai_doaj_org_article_89ec659091f94bb2964188f8e3d92e29 |
| source | Publicly Available Content Database; PubMed Central |
| subjects | Bacteria Bacterial genetics Bacterial Proteins - metabolism Binding proteins Binding sites Caulobacter crescentus Caulobacter crescentus - metabolism chromosome organization chromosome segregation Chromosomes Chromosomes and Gene Expression CTP Cytidine triphosphate Cytidine Triphosphate - metabolism Deoxyribonucleic acid DNA DNA Primase - metabolism DNA, Bacterial - metabolism DNA-binding protein DNA-Binding Proteins - metabolism Hydrolysis Microbiology and Infectious Disease ParB-parS Protein binding Proteins Spreading Transcription Transcription (Genetics) |
| title | ParB spreading on DNA requires cytidine triphosphate in vitro |
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