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J Biol Chem, Vol. 274, Issue 16, 11220-11228, April 16, 1999
From the Division of Hematology/Oncology, Cedars-Sinai Research
Institute/UCLA School of Medicine, Los Angeles, California 90048
Cyclin A1 is a recently cloned cyclin with high
level expression in meiotic cells in the testis. However, it is also
frequently expressed at high levels in acute myeloid leukemia. To
elucidate the regulation of cyclin A1 gene expression, we
cloned and analyzed the genomic structure of cyclin A1. It
consists of 9 exons within 13 kilobase pairs. The TATA-less promoter
initiates transcription from several start sites with the majority of
transcripts beginning within a 4-base pair stretch. A construct
containing a fragment from 
190 to +145 showed the highest
transcriptional activity. Transfection of cyclin A1
promoter constructs into S2 Drosophila cells demonstrated
that Sp1 is essential for the activity of the promoter. Sp1, as well as
Sp3, bound to four GC boxes between nucleotides 130 and 80 as
observed by gel shift analysis. Mutations in two or more of the four GC
boxes decreased promoter activity by >80%. The promoter was found to
be cell cycle-regulated with highest activities found in late S and
G2/M phase. Further analyses suggested that cell cycle
regulation was accomplished by periodic repression of the GC boxes in
G1 phase. Taken together, our data show that cyclin
A1 promoter activity critically depends on four GC boxes, and
members of the Sp1 family appear to be involved in directing expression
of cyclin A1 in both a tissue- and cell cycle-specific manner.
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