UTF1, a novel transcriptional coactivator expressed in pluripotent embryonic stem cells and extra-embryonic cells

Abstract

Corresponding author A.Okuda and A.Fukushima contributed equally to this workWe have obtained a novel transcriptional cofactor, termed undifferentiated embryonic cell transcription factor 1 (UTF1), from F9 embryonic carcinoma (EC) cells. This protein is expressed in EC and embryonic stem cells, as well as in germ line tissues, but could not be detected in any of the other adult mouse tissues tested. Furthermore, when EC cells are induced to differentiate, UTF1 expression is rapidly extinguished. In normal mouse embryos, UTF1 mRNA is present in the inner cell mass, the primitive ectoderm and the extra-embryonic tissues. During the primitive streak stage, the induction of mesodermal cells is accompanied by the down-regulation of UTF1 in the primitive ectoderm. However, its expression is maintained for up to 13.5 days post-coitum in the extra-embryonic tissue. Functionally, UTF1 boosts the level of transcription of the adenovirus E2A promoter. However, unlike the pluripotent cell-specific E1A-like activity, which requires the E2F sites of the E2A promoter for increased transcriptional activation, UTF1-mediated activation is dependent on the upstream ATF site of this promoter. This result indicates that UTF1 is not a major component of the E1A-like activity present in pluripotent embryonic cells. Further analyses revealed that UTF1 interacts not only with the activation domain of ATF-2, but also with the TFIID complex in vivo. Thus, UTF1 displays many of the hallmark characteristics expected for a tissue-specific transcriptional coactivator that works in early embryogenesis.

Negative feedback via RSK modulates Erk‐dependent progression from naïve pluripotency

“…The most differentially regulated phosphopeptides with adj P ‐values of 0.009, 0.017 and 0.019, respectively, were microtubule‐associated protein 1B (Map1B), Jumonji domain containing 1C (Jmjd1c) and p90 ribosomal S6 kinase 1 (RSK1, Rps6ka1 ), all in fraction S1. In N1, increased phosphorylation of pluripotency‐associated transcription factor Utf1 51 was prominent.…”

Differential Requirement for Nucleostemin in Embryonic Stem Cell and Neural Stem Cell Viability

“…As shown in Figure 2A, we confirmed that nucleostemin expression from the Rosa26 locus was rapidly downregulated upon addition of doxycycline to the culture medium. However, the expression levels of pluripotent markers, such as Oct-3/4 [27,32] and UTF1 [25], were unaffected at both the RNA ( Fig. 2A) and protein ( Fig.…”

“…Before performing PCR with the RT products, the number of reaction cycles was determined to achieve semiquantitative conditions with control DNA for each primer set. For western blot analysis, extract treated with sample buffer was used to detect specific protein by chemiluminescence, as described previously [25].…”

“…Generation of anti-Oct-3/4 and undifferentiated embryonic cell transcription factor 1 (UTF1) antibodies were described previously [25,27]. The following commercially available antibodies were used in this study: anti-nucleostemin (AB5691; CHEMI-CON), anti-Fibrillarin (H-140; Santa Cruz Biotechnology, Santa Cruz, CA, http://www.scbt.com), anti-Caspase-3 (9662; Cell Signaling Technology, Beverly, MA, http://www.cellsignal.com), anti-p53 (NCL-p53-CM5p; Novocastra, Newcastle upon Tyne, U.K., http://www.novocastra.…”

Establishment of a new embryonic stem cell line derived from C57BL/6 mouse expressing EGFP ubiquitously

“…We also performed RT-PCR assays to analyze expression of Pou5f1, the activity of which is essential for the identity of the pluripotent cell population in mammalian embryos (Nicols et al, 1998), and Pecam1, Utf1, Cd9, Zfp42, and Spp1, which are expressed in undifferentiated ES cells (Botquin et al, 1998;Hosler et al, 1989;Oka et al, 2002;Okuda et al, 1998;Robson et al, 2001). As shown in Figure 2a, alkaline phosphatase activity was present in colonies composed of ES-like cells on a feeder layer with no activity (Fig.…”