THE UBIQUITOUS TRANSACTIVATOR ZFP-38 IS UP-REGULATED DURING SPERMATOGENESIS WITH DIFFERENTIAL TRANSCRIPTION
THE UBIQUITOUS TRANSACTIVATOR ZFP-38 IS UP-REGULATED DURING SPERMATOGENESIS WITH DIFFERENTIAL TRANSCRIPTION
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DOI：
10.1016/0925-4773(92)90040-q
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发表时间：
1992-12-01
影响因子：
2.6
通讯作者：
FICKENSCHER, H
中科院分区：
文献类型：
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作者：
CHOWDHURY, K;GOULDING, M;FICKENSCHER, H
我们描述了编码一个新的小鼠锌环蛋白基因ZFP-38的全长cDNA克隆，并通过种间回交分析将其定位在5号染色体上。ZFP-38蛋白的N-末端结构域(KDa)由358个氨基酸组成，其C-末端有197个残基，编码7个锌指。我们还提出了ZFP-38是一个强大的转录激活剂的证据。反式激活结构域位于非指状区，含有ZFP-38该区域112个氨基酸残基的融合蛋白和酵母Gal4蛋白的DNA结合区，非常有效地反式激活了带有Gal4靶点的报告CAT质粒的表达。通过原位杂交和Northern印迹技术，可以在精子发生过程中检测到高水平的ZFP-38转录本。它的表达伴随着从粗线期精母细胞到圆形精细胞的发展过程。未分化的精原细胞或单倍体细长的精子细胞和精子不显示任何可检测到的转录本水平。有趣的是，根据核糖核酸酶的保护，其他组织表达低水平的略短的转录本，具有不同的5‘端。ZFP-38的转录激活结构域和7个DNA结合的锌指，再加上细胞类型(S)的特异表达模式，表明ZFP-38在精子发生过程中具有调节转录的潜力。
We describe the complete nucleotide sequence of a full length cDNA clone encoding a new mouse zinc ringer 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.