BOTH LYF-1 AND AN ETS PROTEIN INTERACT WITH A CRITICAL PROMOTER ELEMENT IN THE MURINE TERMINAL TRANSFERASE GENE

被引:60
作者
ERNST, P
HAHM, K
SMALE, ST
机构
[1] UNIV CALIF LOS ANGELES, SCH MED, HOWARD HUGHES MED INST, DEPT MICROBIOL & IMMUNOL, LOS ANGELES, CA 90024 USA
[2] UNIV CALIF LOS ANGELES, SCH MED, INST MOLEC BIOL, LOS ANGELES, CA 90024 USA
关键词
D O I
10.1128/MCB.13.5.2982
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
Terminal deoxynucleotidyltransferase (TdT) is a template-independent DNA polymerase that is expressed transiently during the earliest stages of B- and T-cell ontogeny. Previously, we characterized the promoter for the murine TdT gene and identified a novel DNA-binding protein, called LyF-1, that interacts with a DNA sequence element found to be critical for transcriptional activity in lymphoid cell lines. Here, we present a more detailed analysis of this 30-bp control element, called the TdT D' element, which is centered approximately 60 bp upstream of the transcription start site. We found that both the murine and human D' elements are recognized by multiple proteins, including LyF-1 and at least two Ets family proteins, Ets-1 and Fli-1. Additional protein-DNA interactions were identified through studies using unfractionated nuclear extracts, in which the D' element was apparently incorporated into a multiprotein complex, possibly containing an Ets protein as a core component. By analyzing a series of substitution mutations, two adjacent binding sites for LyF-1 were identified in the murine D' element, with the Ets protein binding site closely coinciding with the proximal, lower-affinity LyF-1 site. Transient transfection analysis with these mutations revealed that only a 10-bp region, containing precisely the Ets and proximal LyF-1 binding sites, was needed for D' activity. These results suggest an important role for an Ets family protein in the expression of the TdT gene. The role of LyF-1 is less clear, it might act in conjunction with the Ets protein bound at the D' element or it might be unnecessary for D' activity.
引用
收藏
页码:2982 / 2992
页数:11
相关论文
共 53 条
[1]   FUNCTIONAL DISSECTION OF THE LCK PROXIMAL PROMOTER [J].
ALLEN, JM ;
FORBUSH, KA ;
PERLMUTTER, RM .
MOLECULAR AND CELLULAR BIOLOGY, 1992, 12 (06) :2758-2768
[2]   DEVELOPMENT OF THE PRIMARY ANTIBODY REPERTOIRE [J].
ALT, FW ;
BLACKWELL, TK ;
YANCOPOULOS, GD .
SCIENCE, 1987, 238 (4830) :1079-1087
[3]   ERYTHROLEUKEMIA INDUCTION BY FRIEND MURINE LEUKEMIA-VIRUS - INSERTIONAL ACTIVATION OF A NEW MEMBER OF THE ETS GENE FAMILY, FLI-1, CLOSELY LINKED TO C-ETS-1 [J].
BENDAVID, Y ;
GIDDENS, EB ;
LETWIN, K ;
BERNSTEIN, A .
GENES & DEVELOPMENT, 1991, 5 (06) :908-918
[4]  
BENOIST C, 1990, ANNU REV IMMUNOL, V8, P681, DOI 10.1146/annurev.iy.08.040190.003341
[5]   THE PRODUCT OF THE C-ETS-1 PROTOONCOGENE AND THE RELATED ETS2 PROTEIN ACT AS TRANSCRIPTIONAL ACTIVATORS OF THE LONG TERMINAL REPEAT OF HUMAN T-CELL LEUKEMIA-VIRUS HTLV-1 [J].
BOSSELUT, R ;
DUVALL, JF ;
GEGONNE, A ;
BAILLY, M ;
HEMAR, A ;
BRADY, J ;
GHYSDAEL, J .
EMBO JOURNAL, 1990, 9 (10) :3137-3144
[7]   CHARACTERIZATION OF SAP-1, A PROTEIN RECRUITED BY SERUM RESPONSE FACTOR TO THE C-FOS SERUM RESPONSE ELEMENT [J].
DALTON, S ;
TREISMAN, R .
CELL, 1992, 68 (03) :597-612
[8]   CHARACTERIZATION OF ANTIGEN RECEPTOR RESPONSE ELEMENTS WITHIN THE INTERLEUKIN-2 ENHANCER [J].
DURAND, DB ;
SHAW, JP ;
BUSH, MR ;
REPLOGLE, RE ;
BELAGAJE, R ;
CRABTREE, GR .
MOLECULAR AND CELLULAR BIOLOGY, 1988, 8 (04) :1715-1724
[9]  
ERNST P, UNPUB
[10]  
FISCHER RJ, 1991, ONCOGENE, V6, P2249