Domain structure of the selenocysteine-specific translation factor SelB in prokaryotes.
Translation factor SelB is the key component for the specific decoding of UGA codons with selenocysteine at the ribosome. SelB binds selenocysteyl-tRNA(Sec), guanine nucleotides and a secondary structure of the selenoprotein mRNA following the UGA at the 3' side. A comparison of the amino acid sequences of SelB species from E. coli, Desulfomicrobium baculatum, Clostridium thermoaceticum and Haemophilus influenzae showed that the proteins consist of at least four structural domains from which the N-terminal three are well conserved and share homology with elongation factor Tu whereas the C-terminal one is more variable and displays no similarity to any protein known. With the aid of the coordinates of EF-Tu the N-terminal part has been modelled into a 3D structure which exhibits intriguing features concerning its interaction with guanine nucleotides and other components of the translational apparatus. Cloning and expression of fragments of SelB and biochemical analysis of the purified truncated proteins showed that the C-terminal 19 kDa protein fragment is able to specifically bind to the selenoprotein mRNA. SelB, thus, is a translation factor functionally homologous to EF-Tu hooked up to the mRNA with its C-terminal end. The formation by SelB of a quaternary complex in vivo has been proven by overexpression of truncated genes of SelB and by demonstration that fragments comprising the mRNA or the tRNA binding domain inhibit selenocysteine insertion.
Böck
A
A
Lehrstuhl für Mikrobiologie, Universität München, Germany.
Hilgenfeld
R
R
Tormay
P
P
Wilting
R
R
Kromayer
M
M
D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D008968:Molecular Conformation; D011387:Prokaryotic Cells; D012333:RNA, Messenger / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry; D016601:RNA-Binding Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism
D001426:Bacterial Proteins; D012333:RNA, Messenger; D012346:RNA, Transfer, Amino Acyl; D016601:RNA-Binding Proteins; C061878:SelB protein, Bacteria; D014157:Transcription Factors; C112705:selenocysteinyl-tRNA