Chowdhury K, Goulding M, Walther C, Imai K, Fickenscher H
Max Planck Institute of Biophysical Chemistry, Department of Molecular Cell Biology, Göttingen, FRG.
Mech Dev. 1992 Dec;39(3):129-42. doi: 10.1016/0925-4773(92)90040-q.
We describe the complete nucleotide sequence of a full length cDNA clone encoding a new mouse zinc finger protein gene, Zfp-38 and localize it on chromosome 5 by the interspecific backcross analysis. The N-terminal domain of the Zfp-38 protein (64 kDa) contains 358 amino acids and the C-terminal domain of 197 residues encodes 7 zinc fingers. We also present evidence that Zfp-38 is a strong transcriptional activator. The transactivation domain was localized in the non finger region and a fusion protein containing 112 amino acid residues from this region of the Zfp-38 and the DNA binding domain of the yeast Gal 4 protein, very efficiently transactivated the expression of a reporter CAT plasmid, harboring the Gal4 target site. By in situ hybridization and northern blotting technique, the Zfp-38 transcript can be detected at a highly elevated level during spermatogenesis. Its expression accompanies the progression from pachytene spermatocytes to round spermatids. The undifferentiated spermatogonia or the haploid elongated spermatid and the spermatozoa do not show any detectable level of the transcript. Interestingly, other tissues express low levels of a slightly shorter transcript with a different 5' end as determined by RNase protection. The presence of both a transcriptional activating domain and 7 DNA binding zinc fingers, coupled with the cell type(s) specific expression pattern, suggests that Zfp-38 has the potential to regulate transcription during spermatogenesis.
我们描述了一个编码新的小鼠锌指蛋白基因Zfp - 38的全长cDNA克隆的完整核苷酸序列,并通过种间回交分析将其定位在5号染色体上。Zfp - 38蛋白(64 kDa)的N端结构域包含358个氨基酸,而197个残基的C端结构域编码7个锌指。我们还提供证据表明Zfp - 38是一种强大的转录激活因子。转录激活结构域定位于非锌指区域,并且一个包含来自Zfp - 38该区域的112个氨基酸残基与酵母Gal 4蛋白的DNA结合结构域的融合蛋白,能非常有效地激活携带Gal4靶位点的报告基因CAT质粒的表达。通过原位杂交和Northern印迹技术,在精子发生过程中可以检测到Zfp - 38转录本的高水平表达。其表达伴随着从粗线期精母细胞到圆形精子细胞的进程。未分化的精原细胞或单倍体延长型精子细胞以及精子均未显示出任何可检测到的转录本水平。有趣的是,通过核糖核酸酶保护分析确定,其他组织表达低水平的、5'端不同的稍短转录本。转录激活结构域和7个DNA结合锌指的存在,再加上细胞类型特异性表达模式,表明Zfp - 38有可能在精子发生过程中调节转录。