TY - JOUR
T1 - Identification of a melanocyte-type promoter of the microphthalmia-associated transcription factor gene
AU - Fuse, Nobuo
AU - Yasumoto, Ken Ichi
AU - Suzuki, Hiroyuki
AU - Takahashi, Kazuhiro
AU - Shibahara, Shigeki
N1 - Funding Information:
We thank S. Subramani for providing pSV2/L, and T. Kasuga for a human melanoma cell line, HMV-II. N.F. is also grateful to Prof. M. Tamai for encouragement. This work was supported in part by Grants-in-aid for Encouragement of Young Scientist (to K-i. Y.), for Developmental Scientific Research (to S.S.) and for Scientific Research on Priority Areas (to S.S.) from the Ministry of Education, Science, and Culture of Japan.
PY - 1996/2/27
Y1 - 1996/2/27
N2 - Microphthalmia-associated transcription factor (MITF), the human homolog of the mouse microphthalmin gene product, regulates melanocyte-specific transcription of the tyrosinase gene that codes for an essential enzyme in melanin biosynthesis. In this study, we have cloned and characterized the human genomic DNA segment containing a melanocyte-type exon and its 5'-flanking region of the MITF gene. A major transcriptional initiation site was assigned by primer extension and S1 nuclease mapping analyses using melanoma RNA. Subsequently, the fusion genes, containing the identified 5'-flanking region upstream from the firefly luciferase gene, were constructed and were introduced into pigmented melanoma cells or HeLa cells which do not express MITF mRNA. Transient expression assays show that the 5'-flanking region of 2.3 kb is able to confer preferential expression of a luciferase gene in pigment cells. These results establish that the MITF gene contains a melanocyte-specific promoter.
AB - Microphthalmia-associated transcription factor (MITF), the human homolog of the mouse microphthalmin gene product, regulates melanocyte-specific transcription of the tyrosinase gene that codes for an essential enzyme in melanin biosynthesis. In this study, we have cloned and characterized the human genomic DNA segment containing a melanocyte-type exon and its 5'-flanking region of the MITF gene. A major transcriptional initiation site was assigned by primer extension and S1 nuclease mapping analyses using melanoma RNA. Subsequently, the fusion genes, containing the identified 5'-flanking region upstream from the firefly luciferase gene, were constructed and were introduced into pigmented melanoma cells or HeLa cells which do not express MITF mRNA. Transient expression assays show that the 5'-flanking region of 2.3 kb is able to confer preferential expression of a luciferase gene in pigment cells. These results establish that the MITF gene contains a melanocyte-specific promoter.
UR - http://www.scopus.com/inward/record.url?scp=0029917487&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0029917487&partnerID=8YFLogxK
U2 - 10.1006/bbrc.1996.0298
DO - 10.1006/bbrc.1996.0298
M3 - Article
C2 - 8645245
AN - SCOPUS:0029917487
SN - 0006-291X
VL - 219
SP - 702
EP - 707
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -