STRUCTURE &EXPRESSION OF ALCOHOL DEHYDROGENASE GENES

Project: Research project

Description

The long term goals of this research are to determine the
structures of mammalian alcohol dehydrogenase (ADH) genes and
the mechanisms which regulate their expression. These studies
will contribute to our understanding of genetic factors underlying
differences among individuals in the metabolic, pharmacologic
and pathological effects of alcohol consumption. This proposal
focuses upon control of ADH expression. Specific Aims 1 through 4 involve analysis of the regulation of
ADH expression in different tissues of the mouse. ADH enzyme
activity, immunoreactive protein, and mRNA will be analyzed in
several tissues of inbred strains of mice with high and low liver
ADH to determine whether the amount of ADH activity is
determined by the steady-state level of its mRNA. In liver and
kidney, the relative ADH activity and mRNA content correspond,
whereas in stomach we have preliminary evidence that they do
not. This suggests an interesting regulatory mechanism in
stomach. The effects of chronic ethanol exposure upon ADH
expression in liver will also be analyzed. The apparent linkage
between a locus controlling ADH expression in liver and
restriction fragment polymorphisms in the Adh-1 locus will be
tested using recombinant inbred strains of mice. ADH-B cDNA
will be isolated. Specific Aims 5 through 7 involve determination of the sequences
that control both tissue-specificity and quantitative expression of
mouse ADH. The Adh-1 genes of mice with high and low liver
ADH will be cloned, and their structures and sequences compared.
The sequences important in the control of tissue specificity of
Adh-1 expression will be analyzed by constructing chimeric genes
with the putative control sequences attached to an easily
assayable reporter gene such as chloramphenicol
acetyltransferase. The expression of these chimeric genes will be
assayed in cultured rodent and human cells of hepatic and non-
hepatic origin. Similar analyses will be carried out to determine
which sequences are involved in quantitative control of ADH by
comparing control regions from high and low activity strains,
again using both human and rodent cells to test whether such
sequences function across species barriers. Specific Aims 8 and 9 involve analysis of the control of ADH
expression in human liver. There are several sized of beta1
mRNA in human livers, which vary in the length of their 3 non-
translated regions. The size distributions of human liver mRNA
from all three Class I ADH loci will be analyzed. The relative
efficiency with which the multiple potential polyadenylation
signals in the human beta1 gene are utilized will be determined
using chimeric genes, and the potential effects upon expression
will be analyzed. Differences in the 3' ends could be a mechanism
for control of the quantity of ADH in liver.
StatusFinished
Effective start/end date7/1/844/30/13

Funding

  • National Institutes of Health
  • National Institutes of Health: $324,964.00
  • National Institutes of Health
  • National Institutes of Health: $248,228.00
  • National Institutes of Health: $304,403.00
  • National Institutes of Health: $290,997.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $298,000.00
  • National Institutes of Health: $298,000.00
  • National Institutes of Health: $328,239.00
  • National Institutes of Health: $332,011.00
  • National Institutes of Health: $298,000.00
  • National Institutes of Health
  • National Institutes of Health: $34,504.00
  • National Institutes of Health: $321,896.00
  • National Institutes of Health: $305,154.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $312,692.00
  • National Institutes of Health: $298,000.00
  • National Institutes of Health: $289,957.00
  • National Institutes of Health
  • National Institutes of Health: $301,556.00
  • National Institutes of Health: $290,582.00
  • National Institutes of Health

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Alcohol Dehydrogenase
Mammals
Oxidoreductases
Genes
Gene Expression
MHC Class I Genes
Alcohol Drinking
Individuality
Transcription Factors
Alcohols
Chromosomes, Human, Pair 4
Vitamin A
Ethanol
Protein Binding
Transfection
Organ Specificity
Liver
Nucleic Acid Regulatory Sequences
Alcoholism
Pharmacology

ASJC

  • Medicine(all)