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
Hauptverfasser: Lin, Hsiu-Hsu Sophia, Khosla, Meenal, Huang, Hao-Jen, Hsu, Duen-Wei, Michaelis, Christine, Weeks, Gerald, Pears, Catherine
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container_issue 1
container_start_page 49
container_title Developmental biology
container_volume 271
creator Lin, Hsiu-Hsu Sophia
Khosla, Meenal
Huang, Hao-Jen
Hsu, Duen-Wei
Michaelis, Christine
Weeks, Gerald
Pears, Catherine
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.
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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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