Selenocysteine synthesis in mammalia: an identity switch from tRNA(Ser) to tRNA(Sec).
The mechanism of selenocysteine insertion into proteins is distinct from all other amino acids in all lines of descent in that it needs specific protein cofactors and a structurally unique tRNA(Sec). It is first aminoacylated with serine and further recognized among all other serylated serine isoacceptors by a selenocysteine synthase and is converted to selenocysteyl-tRNA(Sec). We present here the complete set of identity elements for selenylation of mammalian seryl-tRNA(Sec) and show that the transplantation of these elements into normal serine tRNA allows its selenylation. Four particular structural motifs differentiate eukaryotic tRNA(Sec) from normal tRNA(Ser): the orientation of the extra arm, the short 4 bp T psi C-stem, the extra long 9 bp acceptor-stem and the elongated 6 bp dihydrouridine-stem. Only the last two are essential and only together sufficient for selenocysteine synthesis, whereby the additional base-pairs of the acceptor-stem may be replaced by non-paired nucleotides. Each exchange of the first three structural motifs mentioned above between tRNA(Ser) and tRNA(Sec) resulted in a significant loss of serylation, indicating that the overall composition of particular structure elements is necessary to maintain normal functions of tRNA(Sec). Since we find that all seryl-tRNAs which are selenylated are also substrates for serine phosphorylation we propose that phosphoseryl-tRNA(Sec) is a storage form of seryl-tRNA(Sec).
Institut für Biochemie, Bayerische Julius-Maximilians-Universität, Biozentrum, Würzburg, Germany.
R
R
Amberg
T
T
Mizutani
X Q
XQ
Wu
H J
HJ
Gross
D000215:Acylation; D000818:Animals; D001483:Base Sequence; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008322:Mammals; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010766:Phosphorylation; D017354:Point Mutation; D012345:RNA, Transfer, Amino Acid-Specific / Q000737:chemistry* / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry* / Q000378:metabolism; D012362:RNA, Transfer, Ser / Q000737:chemistry* / Q000378:metabolism; D017279:Selenocysteine / Q000096:biosynthesis* / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D018038:Sodium Selenite / Q000378:metabolism; D014166:Transferases / Q000378:metabolism
D012345:RNA, Transfer, Amino Acid-Specific; D012346:RNA, Transfer, Amino Acyl; D012362:RNA, Transfer, Ser; C036928:tRNA, selenocysteine-; D017279:Selenocysteine; D012694:Serine; D014166:Transferases; C060600:selenium transferase; D018038:Sodium Selenite