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Volume 271, Number 34, Issue of August 23, 1996 pp. 20836-20844
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Genomic Organization and Chromosomal Assignment of the Human 1,4-N-Acetylgalactosaminyltransferase Gene
IDENTIFICATION OF MULTIPLE TRANSCRIPTION UNITS

(Received for publication, December 11, 1995, and in revised form, June 5, 1996)

Keiko Furukawa , Hidenobu Soejima ^¶ , Norio Niikawa ^¶ , Hiroshi Shiku ^§ and Koichi Furukawa

From the Departments of  Oncology and ^¶ Human Genetics, Nagasaki University School of Medicine, 1-12-4 Sakamoto, Nagasaki 852 and the ^§ Department of Internal Medicine II, Mie University School of Medicine, Edohashi, Tsu, Mie 514, Japan

The 1,4-N-acetylgalactosaminyltransferase (1,4GalNAc-T) (EC) gene is expressed in normal brain tissues and in various malignant transformed cells, such as malignant melanoma, neuroblastoma, and adult T cell leukemia. To analyze the regulatory mechanisms of gene expression, we determined the genomic organization of the 1,4GalNAc-T gene. The gene consists of at least 11 exons and spans >8 kilobase pairs. The coding region is located in exons 2-11. To determine the transcription initiation sites, 5-rapid amplification of cDNA ends analysis and ribonuclease protection assays were performed using RNA obtained from the human melanoma cell line SK-MEL-31. Consequently, we defined three transcription initiation sites and the alternative usage of three exons. Exons 1a and 1b partially overlap; the latter is part (3-side) of the former and corresponds to the 5-noncoding region of the cDNA clone previously isolated. The third transcript, exon 1c, corresponds to nucleotides 520 to 412 (position +1 = A of ATG of 1,4GalNAc-T cDNA), which are considered to be in intron 1 based on the cloned cDNA sequence. Ribonuclease protection assays revealed the corresponding protection bands in samples of the gene-expressing cell lines. 5-Flanking regions of individual initiation sites showed promoter activity when analyzed by chloramphenicol acetyltransferase assay in SK-MEL-31 cells. The multiple transcription initiation sites and their promoters/enhancers identified here might be differentially involved in the cell type-specific expression of the 1,4GalNAc-T gene. This gene was assigned to human chromosome 12q13.3 by means of fluorescence in situ hybridization.

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