Negative and positive cis-acting elements in the promoter of the mouse gene that encodes the serine/glycine-rich peptide core of secretory granule proteoglycans.
The gene that encodes a proteoglycan peptide core rich in serine and glycine (SG-PG) is selectively expressed by hematopoietic cells that store in their cytoplasmic granules negatively charged proteoglycans bound ionically to numerous positively charged proteins. With deletion analysis, a negative transcription regulatory element was located between residues -250 and -190 of the 5'-flanking region of the mouse SG-PG gene, and a positive regulatory element was located between residues -118 and -81. The negative regulatory element was dominantly active in fibroblasts that do not express the SG-PG gene whereas the positive regulatory element was dominantly active in hematopoietic cells that do express the SG-PG gene. Site-directed mutagenesis was used to demonstrate that the proximal element within the gene's atypical promoter resided between residues -40 and -20. As assessed by gel mobility shift analyses, the nuclei of rat basophilic leukemia-1 cells and rat-1 fibroblasts contain a number of trans-acting factors that interact with the positive and negative cis-acting regulatory elements of the SG-PG gene. Furthermore, some of these trans-acting factors appear to be different for the two cell types. These studies on cell types that do and do not express the SG-PG gene indicate that transcription of this proteoglycan peptide core gene is regulated constitutively by both positive and negative cis-acting elements located 5' of an atypical promoter.
Avraham
S
S
Department of Medicine, Harvard Medical School, Boston, Massachusetts.
Avraham
H
H
Austen
K F
KF
Stevens
R L
RL
D016475:3T3 Cells; D053674:Aggrecans; D000818:Animals; D001483:Base Sequence; D003594:Cytoplasmic Granules / Q000378:metabolism*; D004247:DNA / Q000235:genetics; D004742:Enhancer Elements, Genetic; D016326:Extracellular Matrix Proteins; D005998:Glycine / Q000235:genetics*; D006023:Glycoproteins / Q000235:genetics*; D013006:Growth Hormone / Q000235:genetics; D037181:Lectins, C-Type; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011509:Proteoglycans; D051381:Rats; D011916:Rats, Inbred F344; D012694:Serine / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
C508320:Acan protein, mouse; C508321:Acan protein, rat; D053674:Aggrecans; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D012694:Serine; D013006:Growth Hormone; D004247:DNA; D005998:Glycine