Assembly of Transcriptionally Inactive Chromatin In Vitro
We have successfully uncoupled the previously interlocked activities of chromatin assembly and in vitro transcription promoted by the Xenopus oocyte S-150 cell-free extract. Our isolated fraction catalyzes extensive chromatin assembly measured both by changes in DNA topology and Micrococcal nuclease...
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| Veröffentlicht in: | Journal of biochemistry (Tokyo) 1989, Vol.106 (1), p.29-33 |
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| container_title | Journal of biochemistry (Tokyo) |
| container_volume | 106 |
| creator | Shanahan, Miya M. Kmiec, Eric B. |
| description | We have successfully uncoupled the previously interlocked activities of chromatin assembly and in vitro transcription promoted by the Xenopus oocyte S-150 cell-free extract. Our isolated fraction catalyzes extensive chromatin assembly measured both by changes in DNA topology and Micrococcal nuclease digestions. The assembly of chromatin is slowed by the exogenous addition of ATP. In the absence of exogenously added ATP, the fraction forms a chromatin template that is transcriptionally inert. Addition of small amounts of the HeLa cell extract (S-100) converts these templates into transcriptionally active ones without disrupting the chromatin structure. Our protocol defines a method for the isolation of a fraction from the Xenopus cell free extract that catalyzes the assembly of transcriptionally inactive chromatin. We characterize this reaction and establish conditions for the transcriptional activation of these inactive minichromosomes. |
| doi_str_mv | 10.1093/oxfordjournals.jbchem.a122813 |
| format | Article |
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Our isolated fraction catalyzes extensive chromatin assembly measured both by changes in DNA topology and Micrococcal nuclease digestions. The assembly of chromatin is slowed by the exogenous addition of ATP. In the absence of exogenously added ATP, the fraction forms a chromatin template that is transcriptionally inert. Addition of small amounts of the HeLa cell extract (S-100) converts these templates into transcriptionally active ones without disrupting the chromatin structure. Our protocol defines a method for the isolation of a fraction from the Xenopus cell free extract that catalyzes the assembly of transcriptionally inactive chromatin. We characterize this reaction and establish conditions for the transcriptional activation of these inactive minichromosomes.</description><identifier>ISSN: 0021-924X</identifier><identifier>EISSN: 1756-2651</identifier><identifier>DOI: 10.1093/oxfordjournals.jbchem.a122813</identifier><identifier>PMID: 2777749</identifier><identifier>CODEN: JOBIAO</identifier><language>eng</language><publisher>Oxford: Oxford University Press</publisher><subject>Animals ; Biological and medical sciences ; Cell-Free System ; Chemical Fractionation ; Chromatin - biosynthesis ; Chromatin - genetics ; Chromatin. Chromosome ; DNA, Circular - metabolism ; DNA, Superhelical - metabolism ; Fundamental and applied biological sciences. 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Our isolated fraction catalyzes extensive chromatin assembly measured both by changes in DNA topology and Micrococcal nuclease digestions. The assembly of chromatin is slowed by the exogenous addition of ATP. In the absence of exogenously added ATP, the fraction forms a chromatin template that is transcriptionally inert. Addition of small amounts of the HeLa cell extract (S-100) converts these templates into transcriptionally active ones without disrupting the chromatin structure. Our protocol defines a method for the isolation of a fraction from the Xenopus cell free extract that catalyzes the assembly of transcriptionally inactive chromatin. We characterize this reaction and establish conditions for the transcriptional activation of these inactive minichromosomes.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cell-Free System</subject><subject>Chemical Fractionation</subject><subject>Chromatin - biosynthesis</subject><subject>Chromatin - genetics</subject><subject>Chromatin. Chromosome</subject><subject>DNA, Circular - metabolism</subject><subject>DNA, Superhelical - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression Regulation</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>RNA, Transfer - genetics</subject><subject>Templates, Genetic</subject><subject>Transcription, Genetic</subject><subject>Xenopus</subject><subject>Xenopus laevis</subject><issn>0021-924X</issn><issn>1756-2651</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEtPwkAUhSdGg4j-BJNucFfsnVc7SyDyMCRukBA3zbSdhsG2gzOtgX9veYStd3Nzz_lycnMQ6kMwgECQV7PPjc22prGVLNxgm6QbVQ4kYBwBuUFdCBn3MWdwi7pBgMEXmK7v0YNz2-OJCemgDg7boaKLxNA5VSbFwTO5t7SycqnVu1qbNrwV55VMa_2rvPHGmlLWumolb6Vrax7RXd4-oJ4uu4c-J2_L8cxffEzn4-HC11iQ2hcqSjAonkvOApwLFkjMQwpYKhHIVEDCM5pSyiiQUIUYBDk6wHOVMRVQ0kMv59ydNT-NcnVcapeqopCVMo2LQwFRFFH2LwiMA8AJfL6ATVKqLN5ZXUp7iC-ltH7_4kuXyiJvW0m1u2IgsMDkxPlnTrta7a--tN8xD0nI4tn6K55NxGg1mr7HC_IHwQ6FWw</recordid><startdate>1989</startdate><enddate>1989</enddate><creator>Shanahan, Miya M.</creator><creator>Kmiec, Eric B.</creator><general>Oxford University Press</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7TM</scope><scope>7X8</scope></search><sort><creationdate>1989</creationdate><title>Assembly of Transcriptionally Inactive Chromatin In Vitro</title><author>Shanahan, Miya M. ; Kmiec, Eric B.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-i293t-9e8b21e6fa6502f950a267412ae90ac91b6d4c4454137e72193ae9016fed5e043</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Cell-Free System</topic><topic>Chemical Fractionation</topic><topic>Chromatin - biosynthesis</topic><topic>Chromatin - genetics</topic><topic>Chromatin. Chromosome</topic><topic>DNA, Circular - metabolism</topic><topic>DNA, Superhelical - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression Regulation</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>RNA, Transfer - genetics</topic><topic>Templates, Genetic</topic><topic>Transcription, Genetic</topic><topic>Xenopus</topic><topic>Xenopus laevis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Shanahan, Miya M.</creatorcontrib><creatorcontrib>Kmiec, Eric B.</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of biochemistry (Tokyo)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Shanahan, Miya M.</au><au>Kmiec, Eric B.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Assembly of Transcriptionally Inactive Chromatin In Vitro</atitle><jtitle>Journal of biochemistry (Tokyo)</jtitle><addtitle>J Biochem</addtitle><date>1989</date><risdate>1989</risdate><volume>106</volume><issue>1</issue><spage>29</spage><epage>33</epage><pages>29-33</pages><issn>0021-924X</issn><eissn>1756-2651</eissn><coden>JOBIAO</coden><abstract>We have successfully uncoupled the previously interlocked activities of chromatin assembly and in vitro transcription promoted by the Xenopus oocyte S-150 cell-free extract. Our isolated fraction catalyzes extensive chromatin assembly measured both by changes in DNA topology and Micrococcal nuclease digestions. The assembly of chromatin is slowed by the exogenous addition of ATP. In the absence of exogenously added ATP, the fraction forms a chromatin template that is transcriptionally inert. Addition of small amounts of the HeLa cell extract (S-100) converts these templates into transcriptionally active ones without disrupting the chromatin structure. Our protocol defines a method for the isolation of a fraction from the Xenopus cell free extract that catalyzes the assembly of transcriptionally inactive chromatin. We characterize this reaction and establish conditions for the transcriptional activation of these inactive minichromosomes.</abstract><cop>Oxford</cop><pub>Oxford University Press</pub><pmid>2777749</pmid><doi>10.1093/oxfordjournals.jbchem.a122813</doi><tpages>5</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Journal of biochemistry (Tokyo), 1989, Vol.106 (1), p.29-33 |
| issn | 0021-924X 1756-2651 |
| language | eng |
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| source | J-STAGE Free; MEDLINE; Free Full-Text Journals in Chemistry; Oxford University Press Journals Digital Archive Legacy |
| subjects | Animals Biological and medical sciences Cell-Free System Chemical Fractionation Chromatin - biosynthesis Chromatin - genetics Chromatin. Chromosome DNA, Circular - metabolism DNA, Superhelical - metabolism Fundamental and applied biological sciences. Psychology Gene Expression Regulation Molecular and cellular biology Molecular genetics RNA, Transfer - genetics Templates, Genetic Transcription, Genetic Xenopus Xenopus laevis |
| title | Assembly of Transcriptionally Inactive Chromatin In Vitro |
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