IDENTIFICATION OF A STAGE SELECTOR ELEMENT IN THE HUMAN GAMMA-GLOBIN GENE PROMOTER THAT FOSTERS PREFERENTIAL INTERACTION WITH THE 5' HS2 ENHANCER WHEN IN COMPETITION WITH THE BETA-PROMOTER

被引:125
作者
JANE, SM
NEY, PA
VANIN, EF
GUMUCIO, DL
NIENHUIS, AW
机构
[1] UNIV MICHIGAN,DEPT ANAT & CELL BIOL,ANN ARBOR,MI 48109
[2] GENET THERAPY INC,GAITHERSBURG,MD 20878
关键词
BETA-GLOBIN; ENHANCER; HYPERSENSITIVITY SITE; PROMOTER; TRANS-ACTING FACTOR;
D O I
10.1002/j.1460-2075.1992.tb05366.x
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 erythroid-specific enhancer within hypersensitivity site 2 (HS2) of the human beta-globin locus control region is required for high level globin gene expression. We investigated interaction between HS2 and the gamma- and beta-promoters using reporter constructs in transient assays in human erythroleukemia (K562) cells. The beta-promoter, usually silent in K562 cells, was activated by HS2. This activity was abolished when a gamma-promoter was linked in cis. Analysis of truncation mutants suggested that sequences conveying the competitive advantage of the gamma-promoter for HS2 included those between positions -53 and -35 relative to the transcriptional start site. This sequence, when used to replace the corresponding region of the beta-promoter, increased beta-promoter activity 10-fold when linked to HS2. The modified beta-promoter was also capable of competing with gamma-promoter modified internally in the -53 to -35 region, when the two promoters were linked to HS2 in a single plasmid. The corresponding sequences from the Galago gamma-promoter, a species which lacks fetal gamma-gene expression, were inactive in analogous assays. We have identified and partially purified a nuclear protein found in human (fetal stage) erythroleukemia cells, but present in much lower concentration in murine (adult stage) erythroleukemia cells, that binds the -53 to -35 sequence of the gamma-promoter. We speculate that this region of the gamma-promoter functions as a stage selector element in the regulation of hemoglobin switching in humans.
引用
收藏
页码:2961 / 2969
页数:9
相关论文
共 50 条
[1]   HUMAN GAMMA-GLOBIN TO BETA-GLOBIN GENE SWITCHING IN TRANSGENIC MICE [J].
BEHRINGER, RR ;
RYAN, TM ;
PALMITER, RD ;
BRINSTER, RL ;
TOWNES, TM .
GENES & DEVELOPMENT, 1990, 4 (03) :380-389
[2]   EMBRYONIC-FETAL ERYTHROID CHARACTERISTICS OF A HUMAN-LEUKEMIC CELL-LINE [J].
BENZ, EJ ;
MURNANE, MJ ;
TONKONOW, BL ;
BERMAN, BW ;
MAZUR, EM ;
CAVALLESCO, C ;
JENKO, T ;
SNYDER, EL ;
FORGET, BG ;
HOFFMAN, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (06) :3509-3513
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   AN EMBRYONIC PATTERN OF EXPRESSION OF A HUMAN-FETAL GLOBIN GENE IN TRANSGENIC MICE [J].
CHADA, K ;
MAGRAM, J ;
COSTANTINI, F .
NATURE, 1986, 319 (6055) :685-689
[5]   SPECIFIC EXPRESSION OF A FOREIGN BETA-GLOBIN GENE IN ERYTHROID-CELLS OF TRANSGENIC MICE [J].
CHADA, K ;
MAGRAM, J ;
RAPHAEL, K ;
RADICE, G ;
LACY, E ;
COSTANTINI, F .
NATURE, 1985, 314 (6009) :377-380
[6]   DEVELOPMENTAL REGULATION OF BETA-GLOBIN GENE SWITCHING [J].
CHOI, ORB ;
ENGEL, JD .
CELL, 1988, 55 (01) :17-26
[7]   FIREFLY LUCIFERASE GENE - STRUCTURE AND EXPRESSION IN MAMMALIAN-CELLS [J].
DEWET, JR ;
WOOD, KV ;
DELUCA, M ;
HELINSKI, DR ;
SUBRAMANI, S .
MOLECULAR AND CELLULAR BIOLOGY, 1987, 7 (02) :725-737
[8]   ACCURATE TRANSCRIPTION INITIATION BY RNA POLYMERASE-II IN A SOLUBLE EXTRACT FROM ISOLATED MAMMALIAN NUCLEI [J].
DIGNAM, JD ;
LEBOVITZ, RM ;
ROEDER, RG .
NUCLEIC ACIDS RESEARCH, 1983, 11 (05) :1475-1489
[9]   HUMAN GAMMA-GLOBIN GENES SILENCED INDEPENDENTLY OF OTHER GENES IN THE BETA-GLOBIN LOCUS [J].
DILLON, N ;
GROSVELD, F .
NATURE, 1991, 350 (6315) :252-254
[10]  
DONOVANPELUSO M, 1987, J BIOL CHEM, V262, P17051