Nilsson M, Bohm S
Center for Nutrition and Toxicology, Karolinska Institute, NOVUM, Huddinge, Sweden.
J Virol. 1994 Jan;68(1):276-88. doi: 10.1128/JVI.68.1.276-288.1994.
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.
小鼠VL30元件构成了逆转录转座子家族的一个成员,在小鼠染色体中以100至200个拷贝的形式分散存在。基于序列同源性,我们已将小鼠VL30成员细分为四个不同的U3亚组。在Northern(RNA)印迹分析中使用亚组特异性探针表明,各个VL30 U3亚组以组织特异性方式表达。我们通过用12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯(TPA)处理的小鼠皮肤进行原位杂交表明,VL30在表皮角质形成细胞中被诱导表达,而在真皮成纤维细胞中不表达。报告基因质粒的瞬时转染以及体外结合分析表明,TPA诱导的角质形成细胞特异性VL30转录由并列位置的两个协同序列基序介导。一个序列基序在角质形成细胞和成纤维细胞中均组成性结合CREB和Jun相关蛋白,而另一个是TPA诱导的c - Rel/p65(NF - κB)结合活性的靶点,且仅在角质形成细胞中存在。这些结合位点在U3亚组以及在TPA处理的小鼠表皮中显示诱导表达的各个U3区域内是保守的。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VL30转录的转录因子的细胞类型特异性活性以及各个U3区域内其同源结合位点的存在与否决定了VL30的诱导性和细胞类型特异性表达。因此,可变的VL30 U3区域可能是研究许多不同细胞系统中诱导性和细胞类型特异性转录的有用工具。