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121.
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123.
Cooperation of proto-signals for nuclear accumulation of estrogen and progesterone receptors. 总被引:35,自引:3,他引:32
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Multiple proto-signals (p-NLSs) for nuclear targeting, none of which suffices on its own, cooperate in the estrogen (ER) and progesterone (PR) receptors. In the ER, an estrogen-inducible p-NLS was found in the hormone binding domain (HBD), in addition to three lysine/arginine-rich motifs resembling prototype constitutive nuclear localization signals (NLSs). The inducible and the constitutive ER p-NLSs cooperate in the presence of estrogen and hydroxy-tamoxifen, but not in the presence of ICI 164,384. In the PR, three p-NLSs, two of which are located within and directly adjacent to the second zinc finger, cooperate with each other and a weak hormone-inducible p-NLS in the PR HBD. No 'masking' of p-NLSs by the HBD was observed for ER and PR, while the ligand-free glucocorticoid receptor HBD inhibited the activity of both homologous and heterologous NLSs. Nuclear co-translocation experiments indicated that in vivo the stability of ER and PR dimers is hormonally controlled, but that, in the absence of the cognate ligand, ER dimers are more stable than PR dimers. This is likely to account for the differential hormone requirement of ER and PR DNA binding in vitro. 相似文献
124.
P. Witzgall M. Bengtsson H. -R. Buser P. -J. Chambon E. Priesner T. Wildbolz H. Arn 《Entomologia Experimentalis et Applicata》1991,60(3):219-223
The sex pheromones of Spilonota ocellana D. & S. and Spilonota laricana Hein. (Lepidoptera: Tortricidae) were identified by chemical analysis and field trapping. Female moths of the two species produce (Z)-8-tetradecenyl acetate, (Z)-8-tetradecen-1-ol and dodecyl acetate in almost the same proportions (98:1:1 and 97:3:1). Males of both species were best attracted to a blend of 10:1 to 1:1 Z8-14Ac:Z8-14OH. This indicates that mating barriers other than sex pheromones exist between sympatric populations. 相似文献
125.
126.
Oligonucleotide-directed mutagenesis by microscale ''shot-gun'' gene synthesis. 总被引:33,自引:10,他引:23
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T Grundstr?m W M Zenke M Wintzerith H W Matthes A Staub P Chambon 《Nucleic acids research》1985,13(9):3305-3316
We describe a rapid and efficient microscale method for in vitro site-directed mutagenesis by gene synthesis. Mutants are constructed by "shot-gun ligation" of overlapping synthetic oligonucleotides yielding double stranded synthetic DNA of more than 120 nucleotides in length. The terminal oligonucleotides of the DNA segment to be synthesized are designed to create sticky ends complementary to unique restriction sites of a polylinker present in an M13 vector. The oligonucleotides are hybridized and ligated to the M13 vector without any purification of the synthetic DNA segment. After cloning, about half of the progeny from such shot-gun ligations contained the predicted sequence demonstrating the efficacy of this method for gene synthesis and its potential for the extensive mutational analysis of genes. 相似文献
127.
128.
The N-terminal part of TIF1, a putative mediator of the ligand-dependent activation function (AF-2) of nuclear receptors, is fused to B-raf in the oncogenic protein T18. 总被引:42,自引:7,他引:35
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B Le Douarin C Zechel J M Garnier Y Lutz L Tora P Pierrat D Heery H Gronemeyer P Chambon R Losson 《The EMBO journal》1995,14(9):2020-2033
129.
Phosphorylation on tyrosine of in vitro synthesized human estrogen receptor activates its hormone binding 总被引:5,自引:0,他引:5
A Migliaccio M Di Domenico S Green A de Falco E L Kajtaniak F Blasi P Chambon F Auricchio 《Molecular endocrinology (Baltimore, Md.)》1989,3(7):1061-1069
Hormone binding controls the activity of estradiol receptor. The in vitro synthesized human receptor binds hormone with high affinity and low efficiency (1-4% of the maximal binding). We now report that phosphorylation on tyrosine of the synthetic receptor by an extensively purified calf uterus kinase increases hormone binding towards maximal levels without change in affinity. This is the first direct demonstration that a newly synthesized hormone receptor acquires ligand binding through phosphorylation. The use of in vitro synthesized proteins as substrates for enzymes which cause functional modifications of proteins is very promising because it is easy to identify the modified domains and residues by using deleted and point mutated proteins. Experiments with two estradiol receptor deletion mutants, one which lacks the N-terminal half of the receptor and binds hormone independently from the N-terminal half of the receptor, the other which lacks the C-terminal half of the receptor and contains the domain required to recognize the estradiol responsive elements, show that tyrosine phosphorylation occurs exclusively within or near the hormone binding domain of the receptor. 相似文献
130.