A homologue of Cdk8 is required for spore cell differentiation in Dictyostelium
The Cdk8 proteins are kinases which phosphorylate the carboxy terminal domain (CTD) of RNA polymerase II (Pol II) as well as some transcription factors and, therefore, are involved in the regulation of transcription. Here, we report that a Cdk8 homologue from Dictyostelium discoideum is localized in...
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Veröffentlicht in: | Developmental biology 2004-07, Vol.271 (1), p.49-58 |
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description | The Cdk8 proteins are kinases which phosphorylate the carboxy terminal domain (CTD) of RNA polymerase II (Pol II) as well as some transcription factors and, therefore, are involved in the regulation of transcription. Here, we report that a Cdk8 homologue from Dictyostelium discoideum is localized in the nucleus where it forms part of a high molecular weight complex that has CTD kinase activity. Insertional mutagenesis was used to abrogate gene function, and analysis of the null strain revealed that the DdCdk8 protein plays an important role in spore formation during late development. As previously reported [Dev. Growth Differ. 44 (2002) 213] Ddcdk8− cells also exhibit impaired aggregation, although we report that the severity of the defect depends upon experimental conditions. When aggregation occurs, Ddcdk8− cells form abnormal terminally differentiated structures within which the Ddcdk8− cells differentiate into stalk cells but fail to form spores, indicating a role for DdCdk8 in cell differentiation. When Ddcdk8 is expressed from its own promoter, the protein is able to rescue both the late developmental defect and the impaired aggregation. However, when expressed from an heterologous promoter, only the impaired aggregation is rescued. This result demonstrates that the defect during late development is not a consequence of impaired aggregation and indicates a direct role for DdCdk8 in spore formation. |
doi_str_mv | 10.1016/j.ydbio.2004.03.020 |
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Here, we report that a Cdk8 homologue from Dictyostelium discoideum is localized in the nucleus where it forms part of a high molecular weight complex that has CTD kinase activity. Insertional mutagenesis was used to abrogate gene function, and analysis of the null strain revealed that the DdCdk8 protein plays an important role in spore formation during late development. As previously reported [Dev. Growth Differ. 44 (2002) 213] Ddcdk8− cells also exhibit impaired aggregation, although we report that the severity of the defect depends upon experimental conditions. When aggregation occurs, Ddcdk8− cells form abnormal terminally differentiated structures within which the Ddcdk8− cells differentiate into stalk cells but fail to form spores, indicating a role for DdCdk8 in cell differentiation. When Ddcdk8 is expressed from its own promoter, the protein is able to rescue both the late developmental defect and the impaired aggregation. However, when expressed from an heterologous promoter, only the impaired aggregation is rescued. This result demonstrates that the defect during late development is not a consequence of impaired aggregation and indicates a direct role for DdCdk8 in spore formation.</description><identifier>ISSN: 0012-1606</identifier><identifier>EISSN: 1095-564X</identifier><identifier>DOI: 10.1016/j.ydbio.2004.03.020</identifier><identifier>PMID: 15196949</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; beta-Galactosidase ; Blotting, Northern ; Chemotaxis - physiology ; Chromatography, Gel ; CTD phosphorylation ; Dictyostelium - enzymology ; Dictyostelium - growth & development ; Dictyostelium discoideum ; Fluorescent Antibody Technique ; Gene Expression Regulation, Developmental ; Gene Library ; Mutagenesis, Insertional ; Phenotype ; Precipitin Tests ; Regulation of transcription ; RNA polymerase II ; Sequence Analysis, DNA ; Spore differentiation ; Spores - growth & development ; Transfection</subject><ispartof>Developmental biology, 2004-07, Vol.271 (1), p.49-58</ispartof><rights>2004 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c462t-76f1f715bac8d9ecd3e004e1fce6d26a009ce3b6c36f2ded9c0425c83c6264513</citedby><cites>FETCH-LOGICAL-c462t-76f1f715bac8d9ecd3e004e1fce6d26a009ce3b6c36f2ded9c0425c83c6264513</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S001216060400209X$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65534</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15196949$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lin, Hsiu-Hsu Sophia</creatorcontrib><creatorcontrib>Khosla, Meenal</creatorcontrib><creatorcontrib>Huang, Hao-Jen</creatorcontrib><creatorcontrib>Hsu, Duen-Wei</creatorcontrib><creatorcontrib>Michaelis, Christine</creatorcontrib><creatorcontrib>Weeks, Gerald</creatorcontrib><creatorcontrib>Pears, Catherine</creatorcontrib><title>A homologue of Cdk8 is required for spore cell differentiation in Dictyostelium</title><title>Developmental biology</title><addtitle>Dev Biol</addtitle><description>The Cdk8 proteins are kinases which phosphorylate the carboxy terminal domain (CTD) of RNA polymerase II (Pol II) as well as some transcription factors and, therefore, are involved in the regulation of transcription. Here, we report that a Cdk8 homologue from Dictyostelium discoideum is localized in the nucleus where it forms part of a high molecular weight complex that has CTD kinase activity. Insertional mutagenesis was used to abrogate gene function, and analysis of the null strain revealed that the DdCdk8 protein plays an important role in spore formation during late development. As previously reported [Dev. Growth Differ. 44 (2002) 213] Ddcdk8− cells also exhibit impaired aggregation, although we report that the severity of the defect depends upon experimental conditions. When aggregation occurs, Ddcdk8− cells form abnormal terminally differentiated structures within which the Ddcdk8− cells differentiate into stalk cells but fail to form spores, indicating a role for DdCdk8 in cell differentiation. When Ddcdk8 is expressed from its own promoter, the protein is able to rescue both the late developmental defect and the impaired aggregation. However, when expressed from an heterologous promoter, only the impaired aggregation is rescued. This result demonstrates that the defect during late development is not a consequence of impaired aggregation and indicates a direct role for DdCdk8 in spore formation.</description><subject>Animals</subject><subject>beta-Galactosidase</subject><subject>Blotting, Northern</subject><subject>Chemotaxis - physiology</subject><subject>Chromatography, Gel</subject><subject>CTD phosphorylation</subject><subject>Dictyostelium - enzymology</subject><subject>Dictyostelium - growth & development</subject><subject>Dictyostelium discoideum</subject><subject>Fluorescent Antibody Technique</subject><subject>Gene Expression Regulation, Developmental</subject><subject>Gene Library</subject><subject>Mutagenesis, Insertional</subject><subject>Phenotype</subject><subject>Precipitin Tests</subject><subject>Regulation of transcription</subject><subject>RNA polymerase II</subject><subject>Sequence Analysis, DNA</subject><subject>Spore differentiation</subject><subject>Spores - growth & development</subject><subject>Transfection</subject><issn>0012-1606</issn><issn>1095-564X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkD1PwzAQhi0EouXjFyAhT2wJ5zhxnYEBlU-pUheQ2KzUPoNLErd2gtR_T0orscF0y_O-d_cQcsEgZcDE9TLdmIXzaQaQp8BTyOCAjBmURVKI_O2QjAFYljABYkROYlwCAJeSH5MRK1gpyrwck_kt_fCNr_17j9RbOjWfkrpIA657F9BQ6wONKx-Qaqxrapy1GLDtXNU531LX0junu42PHdaub87Ika3qiOf7eUpeH-5fpk_JbP74PL2dJToXWZdMhGV2wopFpaUpURuOwxfIrEZhMlEBlBr5QmgubGbQlBryrNCSa5GJvGD8lFztelfBr3uMnWpc3F5Ytej7qJiEvJDDw_-Ck1JKYPkA8h2og48xoFWr4JoqbBQDtRWulupHuNoKV8DVIHxIXe7r-0WD5jezNzwANzsABxtfDoOK2mGr0Qx6daeMd38u-AbXHZLb</recordid><startdate>20040701</startdate><enddate>20040701</enddate><creator>Lin, Hsiu-Hsu Sophia</creator><creator>Khosla, Meenal</creator><creator>Huang, Hao-Jen</creator><creator>Hsu, Duen-Wei</creator><creator>Michaelis, Christine</creator><creator>Weeks, Gerald</creator><creator>Pears, Catherine</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><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>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>20040701</creationdate><title>A homologue of Cdk8 is required for spore cell differentiation in Dictyostelium</title><author>Lin, Hsiu-Hsu Sophia ; Khosla, Meenal ; Huang, Hao-Jen ; Hsu, Duen-Wei ; Michaelis, Christine ; Weeks, Gerald ; Pears, Catherine</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c462t-76f1f715bac8d9ecd3e004e1fce6d26a009ce3b6c36f2ded9c0425c83c6264513</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Animals</topic><topic>beta-Galactosidase</topic><topic>Blotting, Northern</topic><topic>Chemotaxis - physiology</topic><topic>Chromatography, Gel</topic><topic>CTD phosphorylation</topic><topic>Dictyostelium - enzymology</topic><topic>Dictyostelium - growth & development</topic><topic>Dictyostelium discoideum</topic><topic>Fluorescent Antibody Technique</topic><topic>Gene Expression Regulation, Developmental</topic><topic>Gene Library</topic><topic>Mutagenesis, Insertional</topic><topic>Phenotype</topic><topic>Precipitin Tests</topic><topic>Regulation of transcription</topic><topic>RNA polymerase II</topic><topic>Sequence Analysis, DNA</topic><topic>Spore differentiation</topic><topic>Spores - growth & development</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lin, Hsiu-Hsu Sophia</creatorcontrib><creatorcontrib>Khosla, Meenal</creatorcontrib><creatorcontrib>Huang, Hao-Jen</creatorcontrib><creatorcontrib>Hsu, Duen-Wei</creatorcontrib><creatorcontrib>Michaelis, Christine</creatorcontrib><creatorcontrib>Weeks, Gerald</creatorcontrib><creatorcontrib>Pears, Catherine</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Developmental biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lin, Hsiu-Hsu Sophia</au><au>Khosla, Meenal</au><au>Huang, Hao-Jen</au><au>Hsu, Duen-Wei</au><au>Michaelis, Christine</au><au>Weeks, Gerald</au><au>Pears, Catherine</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A homologue of Cdk8 is required for spore cell differentiation in Dictyostelium</atitle><jtitle>Developmental biology</jtitle><addtitle>Dev Biol</addtitle><date>2004-07-01</date><risdate>2004</risdate><volume>271</volume><issue>1</issue><spage>49</spage><epage>58</epage><pages>49-58</pages><issn>0012-1606</issn><eissn>1095-564X</eissn><abstract>The Cdk8 proteins are kinases which phosphorylate the carboxy terminal domain (CTD) of RNA polymerase II (Pol II) as well as some transcription factors and, therefore, are involved in the regulation of transcription. Here, we report that a Cdk8 homologue from Dictyostelium discoideum is localized in the nucleus where it forms part of a high molecular weight complex that has CTD kinase activity. Insertional mutagenesis was used to abrogate gene function, and analysis of the null strain revealed that the DdCdk8 protein plays an important role in spore formation during late development. As previously reported [Dev. Growth Differ. 44 (2002) 213] Ddcdk8− cells also exhibit impaired aggregation, although we report that the severity of the defect depends upon experimental conditions. When aggregation occurs, Ddcdk8− cells form abnormal terminally differentiated structures within which the Ddcdk8− cells differentiate into stalk cells but fail to form spores, indicating a role for DdCdk8 in cell differentiation. When Ddcdk8 is expressed from its own promoter, the protein is able to rescue both the late developmental defect and the impaired aggregation. 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subjects | Animals beta-Galactosidase Blotting, Northern Chemotaxis - physiology Chromatography, Gel CTD phosphorylation Dictyostelium - enzymology Dictyostelium - growth & development Dictyostelium discoideum Fluorescent Antibody Technique Gene Expression Regulation, Developmental Gene Library Mutagenesis, Insertional Phenotype Precipitin Tests Regulation of transcription RNA polymerase II Sequence Analysis, DNA Spore differentiation Spores - growth & development Transfection |
title | A homologue of Cdk8 is required for spore cell differentiation in Dictyostelium |
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