Inducible and cell type-specific expression of VL30 U3 subgroups correlate with their enhancer design

Inducible and cell type-specific expression of VL30 U3 subgroups correlate with their enhancer design
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VL30 U3 亚组的诱导型和细胞类型特异性表达与其增强子设计相关

DOI:
10.1128/jvi.68.1.276-288.1994
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发表时间:
1994
影响因子:
5.4
通讯作者:
S. Bohm
S. Bohm
中科院分区:
医学2区
文献类型:
--
作者:
M. Nilsson;S. Bohm

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鼠VL 30元件构成一个逆转录转座子家族,其以分散在小鼠染色体中的100至200个拷贝表示。基于序列同源性,我们将小鼠VL 30成员细分为四个不同的U3亚组。在北方(RNA)印迹分析中使用亚组特异性探针表明,各个VL 30 U3亚组以组织特异性方式表达。我们表明,通过原位杂交的小鼠皮肤处理与12-O-十四酰基佛波醇-13-乙酸酯(TPA),VL 30的表达诱导表皮角质形成细胞,但不是在真皮成纤维细胞。瞬时转染的报告基因质粒一起在体外结合分析表明,TPA诱导的VL 30转录角质形成细胞特异性介导的两个合作的序列基序并列的位置。一个序列基序显示在角质形成细胞和成纤维细胞中组成性结合CREB-和Jun-相关蛋白,而另一个是TPA诱导的c-Rel/p65(NF-κ B)-结合活性的靶点,特异性地在角质形成细胞中。这些结合位点被发现是保守的U3亚组和个别U3地区显示诱导表达TPA处理的小鼠表皮。这些结果与不同U3亚组之间的序列比较一起表明,已知调节VL 30转录的转录因子的细胞类型特异性活性以及单个U3区域内其同源结合位点的存在或不存在决定了可诱导的和细胞类型特异性的VL 30表达。因此可变VL 30 U3区可能是研究许多不同细胞系统中的诱导型和细胞类型特异性转录的有用工具。
The murine VL30 elements constitute one family of retrotransposons represented in 100 to 200 copies that are dispersed among the mouse chromosomes. On the basis of sequence homology, we have subdivided mouse VL30 members into four distinct U3 subgroups. The use of subgroup-specific probes in Northern (RNA) blot analyses shows that individual VL30 U3 subgroups are expressed in a tissue-specific manner. We show by in situ hybridization of mouse skin treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) that VL30 expression is induced in epidermal keratinocytes but not in dermal fibroblasts. Transient transfections of reporter gene plasmids together with in vitro binding analysis indicate that TPA-induced VL30 transcription specific for keratinocytes is mediated by two cooperating sequence motifs in juxtaposed position. One sequence motif is shown to constitutively bind CREB- and Jun-related proteins in both keratinocytes and fibroblasts, whereas the other is a target for TPA-induced c-Rel/p65(NF-kappa B)-binding activity specifically in keratinocytes. These binding sites are found to be conserved within U3 subgroups and individual U3 regions showing induced expression in TPA-treated mouse epidermis. These results together with a sequence comparison between different U3 subgroups indicate that cell type-specific activity of transcription factors known to regulate VL30 transcription and the presence or absence of their cognate binding sites within individual U3 regions determine inducible and cell type-specific VL30 expression. The variable VL30 U3 regions might thus be useful tools to study inducible and cell type-specific transcription in many different cell systems.
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影响因子: --
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发表时间: 1990
影响因子: 10.5
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DOI: 10.1126/science.1700865
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期刊: SCIENCE
影响因子: 56.9
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