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J. Biol. Chem., Vol. 265, Issue 4, 2306-2310, Feb, 1990
JR Smith, TF Osborne, JL Goldstein and MS Brown
Sterol-dependent regulation of the low density lipoprotein (LDL) receptor
promoter has been localized previously to a 16-base pair sequence,
designated repeat 2, in the 5'-flanking region of the gene. In the current
study, we show that the central 10 nucleotides of repeat 2 are crucial for
the sterol regulatory activity. This sequence includes an octamer,
designated sterol regulatory element 1 (SRE-1), which was identified
previously in the promoter of the gene for 3- hydroxy-3-methylglutaryl
coenzyme A synthase, a sterol-regulated enzyme of cholesterol biosynthesis.
We made a series of single-base substitutions within a 1471-base pair
fragment of the intact LDL receptor promoter, introduced the mutant
plasmids into hamster cells by transfection, and measured mRNA levels in
the absence and presence of sterols. Substitutions within the 10-base pair
sequence in repeat 2 largely prevented the induction of transcription which
occurs in the absence of sterols. None of these point mutations affected
transcription in the presence of sterols. Like an enhancer, the SRE-1 in
repeat 2 functioned in an orientation-independent manner. We interpret
these findings to indicate that the SRE-1 of the LDL receptor promoter is a
conditional positive element that cooperates with other elements to enhance
transcription in the absence of sterols and loses its function in the
presence of sterols.
Identification of nucleotides responsible for enhancer activity of sterol regulatory element in low density lipoprotein receptor gene
Department of Molecular Genetics, University of Texas Southwestern Medical Center, Dallas 75235.
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