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Liver (B-type) phosphofructokinase mRNA. Cloning, structure, and expression.

S C Gehnrich¹, N Gekakis, H S Sul

¹Department of Nutrition, Harvard School of Public Health, Boston, Massachusetts 02115.

The Journal of Biological Chemistry

|August 25, 1988

Summary

Researchers isolated mouse liver phosphofructokinase (PFK) mRNA, sequenced its complementary DNA (cDNA), and analyzed its structure and regulation. This PFK enzyme is crucial for carbohydrate metabolism and responds to diet and hormones.

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Area of Science:

Molecular Biology
Biochemistry
Genetics

Background:

Phosphofructokinase (PFK) is a key regulatory enzyme in glycolysis.
Understanding liver PFK gene expression is vital for metabolic research.

Purpose of the Study:

To clone and characterize mouse liver phosphofructokinase (PFK) mRNA.
To investigate the nutritional and hormonal regulation of liver PFK expression.

Main Methods:

Polysome immunoadsorption to enrich for liver PFK mRNA.
cDNA synthesis and cloning into lambda gt11 expression vector.
Screening, antibody selection, Western blotting, and DNA sequencing.

Main Results:

A full-length mouse liver PFK cDNA clone (2708 nucleotides) was isolated and sequenced.

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The deduced protein sequence shows 68% homology to rabbit muscle PFK, with conserved substrate binding sites.

Liver PFK mRNA levels increased 4-fold upon refeeding starved mice, a response modulated by cAMP and diabetes.

Conclusions:

The study provides a detailed molecular characterization of mouse liver PFK.
Liver PFK expression is tightly regulated by nutritional status and hormonal signals.
This research offers insights into metabolic control mechanisms in the liver.