THE EGR1 PROTEIN CONTAINS A DISCRETE TRANSCRIPTIONAL REGULATORY DOMAIN WHOSE DELETION RESULTS IN A TRUNCATED PROTEIN THAT BLOCKS EGR1-INDUCED TRANSCRIPTION

被引:18
作者
CARMAN, JA [1 ]
MONROE, JG [1 ]
机构
[1] UNIV PENN,DEPT PATHOL & LAB MED,PHILADELPHIA,PA 19104
关键词
D O I
10.1089/dna.1995.14.581
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Egr-1 is a ubiquitous immediate-early gene whose expression is induced by a wide range of different stimuli. A requirement for egr-1 expression has been demonstrated in pathways leading to both proliferation and differentiation, suggesting that egr-1 is a critical intermediary in determining the long-term cellular response to a stimulus. To determine how egr-1 coordinates a cellular response to receptor-mediated stimulation, we have developed a transient cotransfection assay to map functional domains in the EGR1 protein. We localized an activation domain to a serine/threonine/proline-rich region between amino acids 174 and 270. Using this information, we designed a mutant that lacks this activation domain, but retains the DNA-binding domain. When cotransfected into fibroblasts with an EGR1-dependent reporter, this mutant inhibited the transcriptional activity of both endogenous EGR1, as well as exogenously expressed, wild-type EGR1 protein. These data demonstrate that the activation domain of EGR1 is critical for the activity of the protein, and that a mutant lacking this domain can dominantly inhibit wild-type EGR1 function.
引用
收藏
页码:581 / 589
页数:9
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