Selenocysteine's mechanism of incorporation and evolution revealed in cDNAs of three glutathione peroxidases.
The nonsense codon, UGA, has for the first time recently been shown to encode selenocysteine in two proteins, mouse glutathione peroxidase (GSH-Px) (EC 1.11.1.9) and bacterial formate dehydrogenase. A co-translational rather than post-translational selenium-incorporation mechanism has been implicated. Furthermore, high expression levels of GSH-Px have suggested that suppression of termination is efficient and specific. We have isolated and characterized pituitary, kidney and placenta cDNAs for bovine, human and mouse GSH-Px respectively. It is demonstrated that this novel suppression event occurs in diverse tissues, in at least three mammalian species and at the translational step. Surprisingly, GSH-Px is shown to be extramitochondrially encoded, indicating a cytosolic suppression event rather than one utilizing the mitochondria's well-documented extended codon-reading ability. Sequence analysis reveals that a simple proximal contextual pattern responsible for readthrough does not exist. Analysis of predicted secondary structures of mRNAs, however, has revealed a conformation which may be unique to selenocysteine proteins and may prove useful as a tool for artificial incorporation of selenocysteines. A human intron for GSH-Px from an unspliced mRNA has been isolated whose position indicates an ancient, divergent evolutionary relationship with thioredoxin-S2, rather than an independent convergent one.
Mullenbach
G T
GT
Chiron Research Laboratories, Chiron Corporation, Emeryville, CA 94608.
Tabrizi
A
A
Irvine
B D
BD
Bell
G I
GI
Tainer
J A
JA
Hallewell
R A
RA
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003545:Cysteine / Q000031:analogs & derivatives* / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D005979:Glutathione Peroxidase / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D015394:Molecular Structure; D012643:Selenium / Q000378:metabolism*; D017279:Selenocysteine
D017279:Selenocysteine; D004247:DNA; D005979:Glutathione Peroxidase; D012643:Selenium; D003545:Cysteine