カンノ ヒトシ
  菅野 仁
   所属   医学部 医学科(東京女子医科大学病院)
   職種   特任教授
論文種別 原著
言語種別 英語
査読の有無 査読あり
表題 Human HKR isozyme: organization of the hexokinase I gene, the erythroid-specific promoter, and transcription initiation site.
掲載誌名 正式名:Molecular genetics and metabolism
略  称:Mol Genet Metab
ISSNコード:1096-7192(Print)1096-7192(Linking)
巻・号・頁 67(2),pp.118-30
著者・共著者 Murakami K, Kanno H, Miwa S, Piomelli S
担当区分 2nd著者
発行年月 1999/06
概要 We previously described a cDNA for the human HKR isozyme, whose sequence is identical to that of the ubiquitous HKI isozyme, except for a unique 5' end sequence. Screening a human genomic library with a DNA fragment containing an erythroid-specific sequence we found one clone including 5' ends for both HKR and HKI genes. The first HKR exon was located 3 kb 5' of the first HKI exon. These results confirmed that HKR is produced from the HKI gene by alternate promoter and splicing. The HKI gene consisted of 19 exons. All exon-intron boundaries are conserved among the genes for hexokinase and glucokinase. The HKI gene length was estimated at over 67 kb. The initiation site for the HKR was identified by primer extension. Its promoter did not have a canonical TATA box, but an inverted GATA at nt -177 (i.e., 36 nt 5' to the transcription initiation site). In the HKR promoter a DNA fragment spanning nt -275 to nt -107 exhibited erythroid-specific activity. However, this was absent in shorter promoter fragments (nt -206 to -107 or nt -229 to -107). The sequence nt -275 to -229, which appeared critical for the erythroid-specific expression of the HKR gene, contained a consensus motif for Sp-1 and GATA, CCAAT, and GGAA motifs. The electrophoretic mobility shift assay (EMSA) suggested erythroid-specific cooperative protein-protein interaction in this region. Deletion of the GATA sequence as well as reaction with a specific antibody identified GATA-1 as one of the interacting proteins.
DOI 10.1006/mgme.1999.2842
文献番号 10356311