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胚胎癌细胞中逆转录病毒基因表达的决定因素。

Determinants of retrovirus gene expression in embryonal carcinoma cells.

作者信息

Akgün E, Ziegler M, Grez M

机构信息

Abteilung für Zellbiologie, Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie, Hamburg, Federal Republic of Germany.

出版信息

J Virol. 1991 Jan;65(1):382-8. doi: 10.1128/JVI.65.1.382-388.1991.

DOI:10.1128/JVI.65.1.382-388.1991
PMID:1845898
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC240528/
Abstract

The expression of Moloney murine leukemia virus is restricted in embryonal carcinoma (EC) cells. To characterize specific mutations necessary for expression of retroviruses in EC cells, we analyzed the expression of retrovirus mutants and recombinants thereof in EC cell lines F9 and PCC4. DNA sequence comparison and functional studies allowed us to define three point mutations in the enhancer region of the viral mutants at positions -345, -326, and -166 and two point mutations within the 5'-untranslated region of the viral genome at positions +164 and +165 that were essential for retrovirus expression in EC cells. DNA fragments derived from either the wild type or mutant viruses were used to search for sequence-specific DNA-binding factors in nuclear extracts from undifferentiated PCC4 cells. A cellular factor was found to bind strongly to sequences within the enhancer region (-354 to -306) of wild-type viruses but only weakly to sequences derived from mutant viruses. This factor was named ECF-I (for EC cell factor I). Retroviral expression in EC cells correlates with decreased binding affinity for ECF-I.

摘要

莫洛尼鼠白血病病毒的表达在胚胎癌细胞（EC细胞）中受到限制。为了鉴定EC细胞中逆转录病毒表达所需的特定突变，我们分析了逆转录病毒突变体及其重组体在EC细胞系F9和PCC4中的表达。DNA序列比较和功能研究使我们能够确定病毒突变体增强子区域中位于-345、-326和-166位置的三个点突变，以及病毒基因组5'-非翻译区中位于+164和+165位置的两个点突变，这些突变对于逆转录病毒在EC细胞中的表达至关重要。来自野生型或突变病毒的DNA片段用于在未分化的PCC4细胞的核提取物中寻找序列特异性DNA结合因子。发现一种细胞因子与野生型病毒增强子区域（-354至-306）内的序列强烈结合，但与突变病毒衍生的序列结合较弱。这种因子被命名为ECF-I（EC细胞因子I）。EC细胞中的逆转录病毒表达与对ECF-I的结合亲和力降低相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20cc/240528/a67935326275/jvirol00044-0408-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20cc/240528/2241fa6a4ff5/jvirol00044-0406-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20cc/240528/6210513e2d42/jvirol00044-0407-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20cc/240528/c08024dff44f/jvirol00044-0407-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20cc/240528/a67935326275/jvirol00044-0408-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20cc/240528/2241fa6a4ff5/jvirol00044-0406-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20cc/240528/6210513e2d42/jvirol00044-0407-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20cc/240528/c08024dff44f/jvirol00044-0407-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20cc/240528/a67935326275/jvirol00044-0408-a.jpg

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