It permits the diffusion of small molecules and the active transport of larger molecules between the nucleus and cytoplasm. nucleus than mouse embryonic fibroblasts (MEFs) do, and that this inhibition occurs in a multi-step manner. hybridization revealed that ES cell nuclei have significantly less MCMV DNA than MEF nuclei. This appears to be facilitated by the fact that ES cells express less heparan sulfate, 1 integrin, and vimentin, and have LP-211 fewer nuclear pores, than MEF. This may reduce the ability of MCMV to attach to and enter through the cellular membrane, translocate to the nucleus, and cross the nuclear membrane in pluripotent stem cells (ES/induced pluripotent stem cells). The results presented here provide perspective on the relationship between CMV susceptibility and cell differentiation. Introduction In humans, cytomegalovirus (CMV), a member of the herpes virus family, is the most significant infectious source of intrauterine infections that cause congenital anomalies. Intrauterine contamination with human cytomegalovirus (HCMV) is usually thought to be responsible for a variety of abnormalities, depending on the timing of fetal contamination, infectious route, and virulence of the LP-211 computer virus [1]. Differential susceptibility of certain early embryonic cells DRIP78 to HCMV contamination may cause abnormal embryogenesis or organogenesis, resulting in central nervous system defects. Previous studies have demonstrated altered susceptibility to CMV contamination among different cell types, including various types of stem/progenitor cells [2], [3], [4], [5], [6], [7]. This LP-211 can cause abnormal embryogenesis and/or organogenesis, which, in turn, results in congenital anomalies [8]. Studies of human subjects have obvious limitations, but CMVs exhibit strict species specificity, and HCMV therefore cannot be studied directly in any laboratory animal. Thus, general CMV pathogenesis has been examined in mice, using murine CMV (MCMV) [9], [10], and in guinea pigs, using guinea pig CMV[11]. Interestingly, mouse embryos injected with MCMV-infected blastocysts do not express viral genes, suggesting that they are not susceptible to MCMV [12]. Further, mouse embryonic stem (ES) cells are non-permissive to MCMV contamination, and the MCMV immediate-early (IE) promoter is not activated in ES cells from transgenic mice [4]. Human NTera2/D1 embryonic carcinoma cells (NT2) are a useful model in which LP-211 to study the regulatory mechanisms behind major immediate-early (MIE) enhancer/promoter silencing during quiescent HCMV contamination [5], [13], [14]. This is because HCMV replication is usually prevented in embryonic NT2 cells, where viral MIE gene expression is usually blocked, but not in differentiated cells [5], [13], [15], [16]. Trichostatin A (TSA), an inhibitor of histone deacetylases, brings about MIE enhancer/promoter reactivation in quiescently infected NT2 [17], independent of cellular differentiation [18]. Treatment with TSA disrupts heterochromatin nucleation at the MIE enhancer/promoter [18], a process akin to the chromatin disruption that accompanies HCMV reactivation in endogenously-infected dendritic cells [6]. Stimulation of the cyclic AMP (cAMP)/protein kinase A signaling pathway drives cAMP response element (CRE)-dependent MIE enhancer/promoter activation in quiescently infected NT2 cells, thus exposing a potential mode of regulating HCMV reactivation [19]. Whether these mechanisms also regulate CMV contamination in ES cells remains unknown. There are multiple stages to the CMV contamination process. First, the computer virus attaches to the (mammalian) host cell surface via conversation between an envelope component and a LP-211 cellular molecule that serves as a receptor. After attachment, the computer virus must cross the plasma membrane during a phase of its life cycle known as penetration. The viral particle is very large, and no infectious core particle has ever been observed in the nucleus; this suggests that the computer virus is usually disassembled prior to nuclear entry. Finally, viral DNA, or a DNA-protein complex, enters the nucleus. MCMV genes are expressed in 3 sequential phases: immediate-early, early, and late [20]. In this work, we investigated the susceptibility of mouse ES cells to MCMV by comparing each step of the.
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