ISOLATION AND CHARACTERIZATION OF TARGET SEQUENCES OF THE CHICKEN CDXA HOMEOBOX GENE

被引:116
作者
MARGALIT, Y
YARUS, S
SHAPIRA, E
GRUENBAUM, Y
FAINSOD, A
机构
[1] HEBREW UNIV JERUSALEM,HADASSAH MED SCH,DEPT CELLULAR BIOCHEM,IL-91010 JERUSALEM,ISRAEL
[2] HEBREW UNIV JERUSALEM,DEPT GENET,IL-91904 JERUSALEM,ISRAEL
关键词
D O I
10.1093/nar/21.21.4915
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
The DNA binding specificity of the chicken homeodomain protein CDXA was studied. Using a CDXA-glutathione-S-tranferase fusion protein, DNA fragments containing the binding site for this protein were isolated. The sources of DNA were oligonucleotides with random sequence and chicken genomic DNA. The DNA fragments isolated were sequenced and tested in DNA binding assays. Sequencing revealed that most DNA fragments are AT rich which is a common feature of homeodomain binding sites. By electrophoretic mobility shift assays it was shown that the different target sequences isolated bind to the CDXA protein with different affinities. The specific sequences bound by the CDXA protein in the genomic fragments isolated, were determined by DNase I footprinting. From the footprinted sequences, the CDXA consensus binding site was determined. The CDXA protein binds the consensus sequence A, A/T, T, A/T, A, T, A/G. The CAUDAL binding site in the ftz promoter is also included in this consensus sequence. When tested, some of the genomic target sequences were capable of enhancing the transcriptional activity of reporter plasmids when introduced into CDXA expressing cells. This study determined the DNA sequence specificity of the CDXA protein and it also shows that this protein can further activate transcription in cells in culture.
引用
收藏
页码:4915 / 4922
页数:8
相关论文
共 49 条
[1]   THE UPSTREAM REGION OF THE HUMAN HOMEOBOX GENE HOX3D IS A TARGET FOR REGULATION BY RETINOIC ACID AND HOX HOMEOPROTEINS [J].
ARCIONI, L ;
SIMEONE, A ;
GUAZZI, S ;
ZAPPAVIGNA, V ;
BONCINELLI, E ;
MAVILIO, F .
EMBO JOURNAL, 1992, 11 (01) :265-277
[2]   DIFFERENCES AND SIMILARITIES IN DNA-BINDING PREFERENCES OF MYOD AND E2A PROTEIN COMPLEXES REVEALED BY BINDING-SITE SELECTION [J].
BLACKWELL, TK ;
WEINTRAUB, H .
SCIENCE, 1990, 250 (4984) :1104-1110
[3]   ORGANIZER-SPECIFIC HOMEOBOX GENES IN XENOPUS-LAEVIS EMBRYOS [J].
BLUMBERG, B ;
WRIGHT, CVE ;
DEROBERTIS, EM ;
CHO, KWY .
SCIENCE, 1991, 253 (5016) :194-196
[4]   DIFFERENTIAL UTILIZATION OF THE SAME READING FRAME IN A XENOPUS HOMEOBOX GENE ENCODES 2 RELATED PROTEINS SHARING THE SAME DNA-BINDING SPECIFICITY [J].
CHO, KWY ;
GOETZ, J ;
WRIGHT, CVE ;
FRITZ, A ;
HARDWICKE, J ;
DEROBERTIS, EM .
EMBO JOURNAL, 1988, 7 (07) :2139-2149
[5]   THE CAUDAL GENE-PRODUCT IS A DIRECT ACTIVATOR OF FUSHI-TARAZU TRANSCRIPTION DURING DROSOPHILA EMBRYOGENESIS [J].
DEAROLF, CR ;
TOPOL, J ;
PARKER, CS .
NATURE, 1989, 341 (6240) :340-343
[6]   A MOUSE GENE HOMOLOGOUS TO THE DROSOPHILA GENE CAUDAL IS EXPRESSED IN EPITHELIAL-CELLS FROM THE EMBRYONIC INTESTINE [J].
DUPREY, P ;
CHOWDHURY, K ;
DRESSLER, GR ;
BALLING, R ;
SIMON, D ;
GUENET, JL ;
GRUSS, P .
GENES & DEVELOPMENT, 1988, 2 (12A) :1647-1654
[7]  
EDEN FC, 1978, BIOCHEMISTRY-US, V17, P5838, DOI 10.1021/bi00619a035
[8]   CELL-SPECIFIC EXPRESSION OF THE RAT INSULIN GENE - EVIDENCE FOR ROLE OF 2 DISTINCT-5' FLANKING ELEMENTS [J].
EDLUND, T ;
WALKER, MD ;
BARR, PJ ;
RUTTER, WJ .
SCIENCE, 1985, 230 (4728) :912-916
[9]   DIFFERENTIAL DNA-SEQUENCE RECOGNITION IS A DETERMINANT OF SPECIFICITY IN HOMEOTIC GENE-ACTION [J].
EKKER, SC ;
VONKESSLER, DP ;
BEACHY, PA .
EMBO JOURNAL, 1992, 11 (11) :4059-4072
[10]   NONIMMUNOLOGICAL PRECIPITATION OF PROTEIN-DNA COMPLEXES USING GLUTATHIONE-S-TRANSFERASE FUSION PROTEINS [J].
FAINSOD, A ;
MARGALIT, Y ;
HAFFNER, R ;
GRUENBAUM, Y .
NUCLEIC ACIDS RESEARCH, 1991, 19 (14) :4005-4005