Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.3.1.108 (TAT)
 2,389 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

This study aimed to evaluate the pattern of gene expression induced by activin A in mouse Embryonic Stem Cells (ESCs). Mouse ES cells cultured in undifferentiated state by leukemia inhibitory factor and feeder layer cells. Following removing these two anti differentiation factors for 5 days and forming Embryoid Bodies (EBs), the cells divided to 8 equal cells per groups. Differentiation procedure was performed in a two staged protocol; Formed EBs for 4 days (Stage one); expanded differentiated ESCs on gelatin coated dishes for one week (stage two). In the stage one, the media of groups 2-7 contained 10, 30 and 100 ng mL(-1) Activin A. The media in stage two was the same for all groups and contained only Fetal Bovine Serum (FBS). The expression of undifferentiated, ectoderm, mesoderm and endoderm markers were compared with relative RT-PCR method and statistically analyzed. The expression of an undifferentiating marker; Nanog was increased in the Activin A treated groups of stage one. The expression of OCT4 reduced in Activin A treated groups in stage two. In the stage one, the expression of Nodal increased by Activin A. expression of sonic hedgehog (Shh) was suppressed in Activin A treated groups of both stages. In stage two, there were significant decrease for the expression of mesoderm (Brachyury) and Nodal and visceral endoderm (GATA4) markers (p < 0.01). The expression of definitive endoderm markers (PDX1, TAT) showed significantly increased in Activin A treated groups (p < 0.01). Activin A induced differentiation in high concentration by imbalance in undifferentiating markers. Nodal has a dual role, undifferentiating effect and regulation of visceral endoderm towards definitive endoderm. Overexpression of Nanog, alteration in the expression of Nodal and Shh inhibition are three mechanisms for explanation of differentiation induced by activin A in ES cells. These mechanisms induces cascade of gene expression that commits ESCs towards definitive endodermal cells.
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PMID:Kinetics of gene expression during exposure of mouse stem cells to activin A. 1957 65

A comprehensive understanding of the functional network of transcription factors establishing and maintaining pluripotency is key for the development of biomedical applications of stem cells. Nanog plays an important role in early development and is essential to induce natural pluripotency in embryonic stem cells (ESCs). Inducible gain-of-function systems allowing a precise control over time and dosage of Nanog activity would be highly desirable to study its vital role in the establishment and maintenance of pluripotency at molecular level. Here we engineered a recombinant cell permeable version of Nanog by fusing it with the cell penetrating peptide TAT. Nanog-TAT can be readily expressed in and purified from E. coli and binds to a consensus Nanog DNA sequence. At cellular level it enhances proliferation and self-renewal of ESCs in the absence of leukemia inhibitory factor (LIF). Nanog-TAT together with LIF acts synergistically as judged by enhanced clonogenicity and activation of an Oct4-promoter-driven GFP reporter gene. Furthermore Nanog-TAT, in the absence of LIF, promotes pluripotency by inhibiting endodermal specification in a Stat3-independent manner. Our results demonstrate that Nanog protein transduction is an attractive tool allowing control over dose and time of addition to the cells for studying the molecular control of pluripotency without genetic manipulation.
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PMID:Cell-permeant recombinant Nanog protein promotes pluripotency by inhibiting endodermal specification. 2468 18