Structural analysis of the human pro-apoptotic gene Bik: chromosomal localization, genomic organization and localization of promoter sequences.
The human Bik gene codes for a strong pro-apoptotic protein BIK. We have used fluorescent in-situ hybridization to establish the chromosomal localization of the Bik gene to 22q13.3. Genomic clones of the Bik gene were identified from a cosmid library of chromosome 22. Detailed analysis of the Bik gene revealed that it spans a region of about 19kb and comprises of five exons. Sequence analysis indicated that the 5' flanking region of Bik lacks canonical TATA and CAAT boxes but directs transcriptional initiation from a single site. A 1.9kb region containing the promoter elements of the Bik gene was identified and was found to direct expression of the reporter cat gene in transient transfection studies. By mutational analysis, the minimal Bik promoter was localized to a region between -211 to +153. Northern blot analysis showed a ubiquitous expression profile of the Bik mRNA with elevated levels of expression in heart and skeletal muscle.
Verma
S
S
Institute for Molecular Virology, St. Louis University Health Sciences Center, 3681 Park Avenue, St. Louis, MO 63110, USA.
Budarf
M L
ML
Emanuel
B S
BS
Chinnadurai
G
G
D017209:Apoptosis / Q000235:genetics*; D051017:Apoptosis Regulatory Proteins; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D002874:Chromosome Mapping; D002892:Chromosomes, Human, Pair 22 / Q000235:genetics; D004247:DNA / Q000737:chemistry / Q000235:genetics; D005091:Exons; D005260:Female; D015870:Gene Expression; D005796:Genes / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007438:Introns; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011506:Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014018:Tissue Distribution
D051017:Apoptosis Regulatory Proteins; C096820:BIK protein, human; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D011506:Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase