Sp1 binds to the adhesion-molecule-on-glia regulatory element that functions as a positive transcription regulatory element in astrocytes.
We analyzed cis-acting elements regulating the expression of the gene encoding adhesion molecule on glia (AMOG) in primary cultured astrocytes from newborn rat cerebrum and cerebellum. The relative promoter activities among the series of 5' sequential deletion mutants are similar to those observed in B103 (rat neuroblastoma cell line) cells. The previously identified AMRE (AMOG regulatory element) of the GAGGCGGGG sequence functions as a positive regulatory element, not only in B103 cells, but also in astrocytes. Binding factors to the element were identified as Sp1 based on the following observations using nuclear extracts from the astrocytes and B103 cells: (1) The interaction of the factors with AMRE analyzed by DNase I footprinting and methylation interference analyses was similar to that of Sp1; (2) The binding of the factors to AMRE competed with an oligonucleotide containing the authentic Sp1 consensus sequence; (3) Sp1-specific antibody interfered with the formation of the AMRE gel retardation complexes. The functional implications of the factors in AMOG gene regulation are discussed.
Kawakami
K
K
Department of Biology, Jichi Medical School, Tochigi, Japan.
Watanabe
Y
Y
Araki
M
M
Nagano
K
K
D000251:Adenosine Triphosphatases; D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D001483:Base Sequence; D027682:Cation Transport Proteins; D015816:Cell Adhesion Molecules, Neuronal / Q000096:biosynthesis / Q000235:genetics*; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology; D002540:Cerebral Cortex / Q000166:cytology; D004274:DNA, Recombinant / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis / Q000235:genetics*; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D010957:Plasmids; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D017384:Sequence Deletion; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D014407:Tumor Cells, Cultured
C504192:Atp1b2 protein, rat; D027682:Cation Transport Proteins; D015816:Cell Adhesion Molecules, Neuronal; D004274:DNA, Recombinant; D016326:Extracellular Matrix Proteins; D011993:Recombinant Fusion Proteins; D016329:Sp1 Transcription Factor; D000251:Adenosine Triphosphatases