THE GENE ENCODING RAT PHOSPHOGLYCERATE MUTASE SUBUNIT-M - CLONING AND PROMOTER ANALYSIS IN SKELETAL-MUSCLE CELLS

被引:13
作者
RUIZLOZANO, P
DELECEA, L
BUESA, C
DELAOSA, PP
LEPAGE, D
GUALBERTO, A
WALSH, K
PONS, G
机构
[1] UNIV BARCELONA, FAC MED, DEPT CIENCIAS FISIOL HUMANES & NUTR, BIOQUIM UNITAT, E-08028 BARCELONA, SPAIN
[2] CASE WESTERN RESERVE UNIV, SCH MED, DEPT PHYSIOL & BIOPHYS, CLEVELAND, OH 44106 USA
[3] UNIV BARCELONA, FAC BIOL, DEPT BIOL CELLULAR, E-08028 BARCELONA, SPAIN
[4] UNIV BARCELONA, FAC FARM, DEPT CIENCIAS FISIOL, BIOQUIM UNITAT, E-08028 BARCELONA, SPAIN
关键词
RECOMBINANT DNA; NUCLEOTIDE SEQUENCE; AMINO-ACID SEQUENCE; EXON; INTRON; RAT; PHAGE LIBRARY; TRANSCRIPTIONAL REGULATORY ELEMENTS;
D O I
10.1016/0378-1119(94)90074-4
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The expression of the gene encoding the muscle-specific (M)-subunit of phosphoglycerate mutase (PGAM-M) is restricted to adult skeletal and cardiac muscle. In order to study its expression in muscle, the rat PGAM-M gene has been isolated and sequenced. Rat PGAM-M spans about 2.2 kb and is composed of three exons: 442, 181 and 186-bp long, and two introns of 97 bp and 1.3 kb. The analysis of the 5'-flanking region reveals a promoter which contains multiple DNA regulatory elements and constitutes an ideal model to study muscle gene transcriptional regulation. Thus, the elements responsible for rat PGAM-M muscle-specific expression have been identified by transient transfection in chicken embryo primary cultures, using chimeric constructs of the rat promoter linked to a cat reporter gene. Here, we report that in spite of the abundance of E-box motifs in the rat PGAM-M promoter, known for their involvement in muscle gene expression, two DNA elements regulate the muscle-specific transcription of rat PGAM-M: an A/T motif, the putative MEF-2-binding site (myocyte-specific enhancer-binding factor 2), and a proximal 27-bp element which is conserved between the rat and human genes. These two elements define a small promoter (170 bp) sufficient to support potent and skeletal-muscle-specific expression. The conserved 27-bp region constitutes a transcriptional regulatory element able to confer muscle-specific expression when located upstream from a heterologous TATA box.
引用
收藏
页码:243 / 248
页数:6
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