Characterization of the promoter region of the c-yes proto-oncogene: the importance of the GC boxes on its promoter activity.
In this study we cloned the 5' flanking sequence of the human c-yes gene and identified its promoter region. A 0.53 kilobase pair (kbp) fragment containing the 5' terminus of the c-yes gene showed strong promoter activity when placed upstream of the bacterial chloramphenicol acetyltransferase (CAT) gene and transfected into monkey CV-1 cells. By nuclease S1 mapping multiple transcriptional start sites were detected within the promoter region. Nucleotide sequence analysis revealed that the c-yes promoter region had high G + C contents (64%) and contained six GC box-like sequences (one at the 5' distal region and five in a cluster at the 5' proximal region), but not a TATA box. These features of the c-yes promoter region are similar to those of other protooncogenes, ras-family genes and c-raf-1, and some house-keeping genes. Deletion analysis suggested that the most downstream 0.21 kbp region is primarily important for the promoter activity. This 0.21 kbp region contains one major and another minor transcriptional start site. Five GC box-like sequences were located within this region, and four of them were shown to bind with purified Sp1 transcription factor. Furthermore, using the base-substituted mutants of the Sp1-binding sites, each GC box in the cluster (GC1 to GC4) was shown to affect the c-yes gene expression.
Department of Oncology, University of Tokyo, Japan.
Y
Y
Matsuzawa
K
K
Semba
J
J
Kawamura-Tsuzuku
T
T
Sudo
S
S
Ishii
K
K
Toyoshima
T
T
Yamamoto
D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004742:Enhancer Elements, Genetic; D005260:Female; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010920:Placenta / Q000502:physiology; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D051575:Proto-Oncogene Proteins c-yes; D011519:Proto-Oncogenes; D015183:Restriction Mapping; D014162:Transfection; D019061:src-Family Kinases
C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins; D015500:Chloramphenicol O-Acetyltransferase; D011505:Protein-Tyrosine Kinases; D051575:Proto-Oncogene Proteins c-yes; D019061:src-Family Kinases