Molecular cloning and analysis of the mouse gicerin gene.
Gicerin is a cell adhesion molecule, which has five immunoglobulin-like loop structures in an extracellular domain followed by a single transmembrane domain and a short cytoplasmic tail. We have reported that gicerin participates in neurite extension and structural organization of the nervous system, and its expression in the nervous system is high during the development and dramatically decreased after birth. To elucidate the mechanism how the expression of gicerin is regulated, we performed a genomic cloning of a mouse gicerin. A fragment of 16 kbp genomic clone contained 8 kbp gicerin gene composed of 16 exons with 6 kbp upstream region. Genomic cloning revealed that two isoforms of gicerin were generated by an alternative splicing of exon 15 results in cytoplasmic domains composed of either 63 or 21 amino acids. As for an expressional regulation of gicerin, we found that the mRNA content of gicerin in PC12 cells was regulated by cAMP. Quantitative-PCR analysis revealed that forskolin induced four-fold increase of gicerin mRNA. To characterize the involvement of its promoter region, we examined the promoter activity in PC12 cells by a luciferase-reporter assay. We found that a CRE site located at 60 bp upstream of gicerin gene was responsible for the increase of its mRNA induced by forskolin.
Kohama
Keiko
K
Department of Pharmacology, Osaka University Medical School, Room# A6, 2-2 Yamada-oka Suita, Osaka 565-0871, Japan.
Tsukamoto
Yasuhiro
Y
Furuya
Masaru
M
Okamura
Koh
K
Tanaka
Hidekazu
H
Miki
Naomasa
N
Taira
Eiichi
E
D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D051929:CD146 Antigen; D003001:Cloning, Molecular; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D005091:Exons / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D005796:Genes / Q000235:genetics*; D005809:Genes, Regulator / Q000187:drug effects / Q000235:genetics; D017930:Genes, Reporter; D019426:Integrases / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D019006:Neural Cell Adhesion Molecules / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D016716:PC12 Cells; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
D015703:Antigens, CD; D051929:CD146 Antigen; D019006:Neural Cell Adhesion Molecules; D012333:RNA, Messenger; D005576:Colforsin; D000242:Cyclic AMP; C045073:Cre recombinase; D019426:Integrases