Gene dosage and Down's syndrome: metabolic and enzymatic changes in PC12 cells overexpressing transfected human liver-type phosphofructokinase.
Down's syndrome (DS) is a human genetic disease caused by triplication of the distal third of chromosome 21 and overexpression of an unknown number of genes residing in it. The gene for the liver-type subunit of phosphofructokinase (PFKL), a key glycolytic enzyme, maps to this region and the product is overproduced in DS erythrocytes and fibroblasts. These facts, together with abnormalities which occur in DS glycolysis, make PFKL overexpression a candidate for causing some aspects of the DS phenotype. A cellular model for examining the consequences of PFKL overexpression in DS was constructed by transfecting rat PC12 cells with the human PFKL cDNA. Phosphofructokinase (PFK) isolated from PFKL-overexpressing clones was more inhibited by ATP and citrate and less activated by fructose-6-phosphate than control PFK; similar results were obtained when PFK preparations from DS and control fibroblasts were compared. In vivo NMR measurements determined that cells overexpressing PFKL performed glycolysis 40% faster than controls. These results show that overexpression of PFKL is the cause for altered biochemical regulatory characteristics of PFK in DS fibroblasts and can result in enhancement of glycolysis rates. It is also shown that increased gene dosage can exert its influence not merely by enhancing the amounts of gene products but also by altering their biochemical nature.
Elson
A
A
Department of Molecular Genetics and Virology, Weizmann Institute of Science, Rehovot, Israel.
Bernstein
Y
Y
Degani
H
H
Levanon
D
D
Ben-Hur
H
H
Groner
Y
Y
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D002951:Citrates / Q000494:pharmacology; D019343:Citric Acid; D003001:Cloning, Molecular; D004314:Down Syndrome / Q000201:enzymology* / Q000235:genetics; D005636:Fructosephosphates / Q000494:pharmacology; D005708:Gallium / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000235:genetics*; D006019:Glycolysis / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D016716:PC12 Cells; D010732:Phosphofructokinase-1 / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D014162:Transfection / Q000235:genetics
D002951:Citrates; D005636:Fructosephosphates; D019343:Citric Acid; C027618:fructose-6-phosphate; D000255:Adenosine Triphosphate; D005708:Gallium; D010732:Phosphofructokinase-1