Project description:Transcriptional profiling of human mesenchymal stem cells comparing normoxic MSCs cells with hypoxic MSCs cells. Hypoxia may inhibit senescence of MSCs during expansion. Goal was to determine the effects of hypoxia on global MSCs gene expression.
Project description:The polyadenosine RNA binding proteins (Pabs) represent one class of RNA binding proteins that play critical roles in gene expression. This class includes the well-studied nuclear and cytoplasmic Pabs, PABPN1 and PABPC1, respectively, as well as the newly characterized nuclear Pab, zinc finger CCCH-type containing #14, or ZC3H14. ZC3H14 was recently linked to a form of intellectual disability, suggesting a critical role for ZC3H14 in neurons; however, the post-transcriptional function of ZC3H14 is unknown. In this study, we performed a microarray analysis of cells depleted of ZC3H14 or PABPN1 in MCF-7 breast cancer cells. These results revealed that PABPN1 significantly affected ~17% of expressed transcripts as compared to ZC3H14, which affected ~1% of expressed transcripts, suggesting that ZC3H14 has specific mRNA targets. The differentially expressed mRNAs identified in this analysis not only provide information about the classes and types of transcripts that are regulated by these proteins, but also represent a set of transcripts that could be directly bound by ZC3H14 and/or PABPN1. Total RNA isolated from MCF-7 cells treated for 48 hours with siRNA targeting PABPN1, ZC3H14 (all splice variants), or a control scramble siRNA.
Project description:During zygote genome activation (ZGA) in embryos, large amounts of precursor mRNA undergo complex post-transcriptional regulation such as splicing in nuclear speckles, and are eventually transported into the cytoplasm for translation into proteins. However, it is unclear whether the nuclear speckles play an important role in embryos. In this study, we demonstrated that ZC3H14 promotes the formation of nuclear speckles, which is essential for mouse embryonic development. Zc3h14 knockout mice had reduced fertility and embryonic development was blocked at the 2-cell stage. ZC3H14 is localized to nuclear speckles during embryonic transcriptional activity, and inhibition of its phosphorylation affects the binding of ZC3H14 to mRNA and inhibits the formation of nuclear speckles. As a result, the mRNA cannot complete alternative splicing normally, and a large amount of abnormal RNA accumulates in the embryo, leading to developmental arrest. Taken together, our results demonstrate that ZC3H14 has an important function in fertility in female mice by promoting nuclear speckles formation.