Functional analysis of the Cdk7.cyclin H.Mat1 complex in mouse embryonic stem cells and embryos.
The trimeric Cdk7.cyclin H.Mat1 complex functions in cell cycle regulation, as the Cdk-activating kinase, and in transcription, as a module of the general transcription factor TFIIH. As a component of TFIIH, Cdk7 phosphorylates serines 5 and 7 of the carboxyl-terminal domain of RNA polymerase II and can also directly phosphorylate transcription factors to regulate gene expression. Here we have investigated the function of the Cdk7.cyclin H.Mat1 complex in murine embryonic stem (ES) cells and preimplantation embryos to determine whether it regulates the unique cell cycle structure and transcriptional network of pluripotent cells. We demonstrate that depletion of cyclin H leads to differentiation of ES cells independent of changes in cell cycle progression. In contrast, we observed that developmental genes are acutely up-regulated after cyclin H down-regulation, likely perturbing normal ES self-renewal pathways. We further demonstrate that Spt5, a known phosphorylation target of Cdk7, similarly regulates ES pluripotency and gene expression. Consistent with its function in ES cells, cyclin H depletion from mouse embryos also leads to defects in the expansion of the inner cell mass of blastocysts, a transient pluripotent stem cell population in vivo. Our findings indicate that cyclin H has an essential function in promoting the self-renewal of the pluripotent stem cells of blastocyst stage embryos. Collectively, these studies demonstrate a critical and novel role for cyclin H in maintaining ES cell identity and suggest that cyclin H has important functions in early embryonic development.
Patel
Shetal A
SA
Abramson Family Cancer Research Institute, University of Pennsylvania, Philadelphia, Pennsylvania 19104; School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104.
Simon
M Celeste
MC
Abramson Family Cancer Research Institute, University of Pennsylvania, Philadelphia, Pennsylvania 19104; School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104; Howard Hughes Medical Institute, University of Pennsylvania, Philadelphia, Pennsylvania 19104. Electronic address: celeste2@mail.med.upenn.edu.
D026921:Amino Acid Transport Systems, Neutral / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D015153:Blotting, Western; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D002454:Cell Differentiation; D002460:Cell Line; D056748:Cyclin H / Q000378:metabolism / Q000502:physiology*; D018844:Cyclin-Dependent Kinases / Q000378:metabolism / Q000502:physiology*; D015536:Down-Regulation; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors; D015854:Up-Regulation
D026921:Amino Acid Transport Systems, Neutral; D018797:Cell Cycle Proteins; D056748:Cyclin H; D018844:Cyclin-Dependent Kinases; D014157:Transcription Factors; C534752:Ccnh protein, mouse; C432410:Mnat1 protein, mouse; C090586:Cdk7 protein, mouse