Ferrochelatase structural mutant (Fechm1Pas) in the house mouse.
The molecular basis of an inherited defect of ferrochelatase in mouse (Fechm1Pas/Fechm1Pas, described by Tutois et al., 1991, J. Clin. Invest. 88:1730-1736) was investigated. cDNA clones encoding ferrochelatase, isolated by amplification of the mRNA from the liver of a mutant mouse using the polymerase chain reaction, were sequenced by the dideoxynucleotide chain-termination method. All the clones carried a T to A transversion at nucleotide 293, leading to a methionine to lysine substitution at position 98 in the protein (mutation M98K). Hybridization with allele-specific oligonucleotides (ASOs) confirmed the mutation at the cDNA and genomic levels. Finally, expression of the mutant ferrochelatase protein in E. coli demonstrated a marked deficiency in activity in agreement with the activity of the deficient enzyme in vivo. This Fechm1Pas/Fechm1Pas mutant mouse represents a useful model for studying the pathophysiological feature of the human disease and the first accessible model for gene therapy in the field of porphyrias.
Boulechfar
S
S
Laboratoire de Génétique Moléculaire, Faculté X. Bichat, Université Paris 7, France.
Lamoril
J
J
Montagutelli
X
X
Guenet
J L
JL
Deybach
J C
JC
Nordmann
Y
Y
Dailey
H
H
Grandchamp
B
B
de Verneuil
H
H
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA; D004926:Escherichia coli; D005294:Ferrochelatase / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D046351:Protoporphyria, Erythropoietic
D004247:DNA; D005294:Ferrochelatase