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J Biol Chem, Vol. 274, Issue 12, 7816-7824, March 19, 1999
From the Department of Food Science and Human Nutrition and the
Division of Nutritional Sciences, University of Illinois,
Urbana, Illinois 61801
We previously showed that rat liver
betaine-homocysteine methyltransferase (BHMT) mRNA content and
activity increased 4-fold when rats were fed a methionine-deficient
diet containing adequate choline, compared with rats fed the same diet
with control levels of methionine (Park, E. I., Renduchintala,
M. S., and Garrow, T. A. (1997) J. Nutr. Biochem.
8, 541-545). A further 2-fold increase was observed in rats fed the
methionine-deficient diet with supplemental betaine. The nutrition
studies reported here were designed to determine whether other methyl
donors would induce rat liver BHMT gene expression when added to a
methionine-deficient diet and to define the relationship between the
degree of methionine restriction and level of methyl donor intake on
BHMT expression. Therefore, rats were fed amino acid-defined diets
varying in methionine and methyl donor composition. The effect of diet
on BHMT expression was evaluated using Northern, Western, and enzyme
activity analyses. Similar to when betaine was added to a
methionine-deficient diet, choline or sulfonium analogs of betaine
induced BHMT expression. The diet-induced induction of hepatic BHMT
activity was mediated by increases in the steady-state level of its
mRNA and immunodetectable protein. Using methyl donor-free diets,
we found that methionine restriction was required but alone not
sufficient for the high induction of BHMT expression. Concomitant with
methionine restriction, dietary methyl groups were required for high
levels of BHMT induction, and a dose-dependent relationship
was observed between methyl donor intake and BHMT induction.
Furthermore, the severity of methionine restriction influenced the
magnitude of BHMT induction.
To study the molecular mechanisms that regulate the expression of BHMT,
we have cloned the human BHMT gene. This gene spans about 20 kilobases
of DNA and contains 8 exons and 7 introns. Using RNA isolated from
human liver and hepatoma cells, a major transcriptional start site has
been mapped using the 5' rapid amplification of cDNA ends
technique, and this start site is 26 nucleotides downstream from a
putative TATA box.
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