The Role of Transcriptional Corepressor Nif3l1 in Early Stage of Neural Differentiation via Cooperation with Trip15/CSN2

Abstract

Mouse Nif3l1 gene is highly conserved from bacteria to human. Even though this gene is expressed throughout embryonic development, its biological function is still obscure. Here, we show that Nif3l1 participates in retinoic acid-primed neural differentiation of P19 embryonic carcinoma cells through cooperation with Trip15/CSN2, a transcriptional corepressor/component of COP9 signalosome. We isolated Nif3l1 cDNA from P19 cell cDNA library by a yeast two-hybrid screening using Trip15/CSN2 as a bait. This interaction was confirmed by a pull-down assay and an epitope-tagged coimmunoprecipitation. Although Nif3l1 was mainly detected in the cytoplasm, the translocation of Nif3l1 into the nuclei was observed in retinoic acid-primed neural differentiation of P19 cells and enhanced by the enforced expression of Trip15/CSN2. Furthermore, enforced expression of sense Nif3l1 RNA, but not antisense RNA, enhanced the neural differentiation of P19 cells accompanying the intense down-regulation of Oct-3/4 mRNA expression and the rapid induction of Mash-1 mRNA expression. Luciferase reporter assay showed that Nif3l1 could act as a transcriptional repressor and synergized the transcriptional repression by Trip15/CSN2. These results indicate that Nif3l1 implicates in neural differentiation through the cooperation with Trip15/CSN2.

The subcellular localization of the ChoRE-binding protein, encoded by the Williams–Beuren syndrome critical region gene 14, is regulated by 14-3-3

“…As previously reported, immunofluorescence microscopy shows that Wbscr14, 14-3-3 and NIF3L1-a localize in the cytoplasm (5,26,28,29). Similarly, Myc -EGFP -NIF3L1-b and Myc -EGFP -NIF3L1-a -L244X show staining in the cytoplasm of COS-7 cells (Supplementary Material, Fig.…”

“…Moreover, loss of yADA3 or of its partner yADA2 affects transcription of a large number of genes and inhibits the rapid global increase in transcription that occurs in response to glucose, as yeast cells resume rapid growth and accelerate progress through the cell cycle (56). The mouse Nif3l1 protein was shown recently to interact with Trip15/CSN2, a transcriptional corepressor/component of the COP9 signalosome thereby executing a crucial role in retinoic acid-primed neural differentiation of P19 embryonic carcinoma cells (28). Interestingly, the Wbscr14-interacting domain maps to the first 243 residues, whereas the Trip15/CSN2-binding moiety was shown to map to the COOH-terminus (residues 243 -376) (this work and 28).…”

Involvement of the Mouse Prp19 Gene in Neuronal/Astroglial Cell Fate Decisions

“…To induce neural differentiation, 1 ϫ 10 6 aggregated P19 cells were cultured in 10-cm bacteriological grade dishes in 10 ml of ␣-minimal essential medium containing 10% fetal calf serum and 5 ϫ 10 Ϫ7 M all-trans-RA (Sigma) for 4 days as described previously (16). The cell aggregates were suspended by mild pipetting and transferred to tissue culture dishes.…”

“…Subtractive Cloning-Tester poly(A) ϩ RNA was prepared from aggregated P19 cells treated with 5 ϫ 10 Ϫ7 M RA for 24 h, and firststrand cDNA was hybridized with excess driver RNA prepared from untreated P19 cells, and then a subtractive cDNA probe was prepared by the addition of the cross-linking agent 2,5-diaziridinyl-1,4-benzoquinone and used to screen 9 ϫ 10 5 plaques of rat brain 5Ј-stretch plus a gt11 cDNA library (Clontech) as described previously (16). The cDNAs inserted into the positive phage were then amplified by PCR using primers based on the phage DNA sequence: 5Ј-primer, 5Ј-GCC ACG ACT CCT GGA GCC CG-3Ј; and 3Ј-primer, 5Ј-CAC CAG ACC AAC TGG TAA TG-3Ј.…”

“…In this context, to clarify the molecular mechanisms in neuronal/ astroglial fate decisions in mammals, we applied a subtractive cDNA cloning strategy for the retinoic acid (RA)-induced neural differentiation system of mouse P19 embryonic carcinoma cells as described previously (16). By this approach, we isolated PRP19␣ (precursor RNA processing-19␣) and its splice variant PRP19␤, with an additional 19-amino acid residues in-frame.…”