TY - JOUR
T1 - A homologue of Cdk8 is required for spore cell differentiation in Dictyostelium
AU - Lin, Hsiu Hsu Sophia
AU - Khosla, Meenal
AU - Huang, Hao Jen
AU - Hsu, Duen Wei
AU - Michaelis, Christine
AU - Weeks, Gerald
AU - Pears, Catherine
N1 - Funding Information:
Many thanks are due to Sasha Akoulitchev for helpful discussions and for reading the manuscript and to Emma Lees for providing the CTD peptide. This work was funded by the BBSRC and The Wellcome Trust (grant no. 063612/Z) (C.P.) and the National Science and Engineering Council of Canada (G.W.). H.H.L. is the grateful recipient of a Swire Taiwan Graduate Scholarship to University College, Oxford.
PY - 2004/7/1
Y1 - 2004/7/1
N2 - 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.
AB - 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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U2 - 10.1016/j.ydbio.2004.03.020
DO - 10.1016/j.ydbio.2004.03.020
M3 - Article
C2 - 15196949
AN - SCOPUS:2942513050
VL - 271
SP - 49
EP - 58
JO - Developmental Biology
JF - Developmental Biology
SN - 0012-1606
IS - 1
ER -