Function of the SmpB tail in transfer-messenger RNA translation revealed by a nucleus-encoded form.
Stalled bacterial ribosomes are freed when they switch to the translation of transfer-messenger RNA (tmRNA). This process requires the tmRNA-binding and ribosome-binding cofactor SmpB, a beta-barrel protein with a protruding C-terminal tail of unresolved structure. Some plastid genomes encode tmRNA, but smpB genes have only been reported from bacteria. Here we identify smpB in the nuclear genomes of both a diatom and a red alga encoding a signal for import into the plastid, where mature SmpB could activate tmRNA. Diatom SmpB was active for tmRNA translation with bacterial components in vivo and in vitro, although less so than Escherichia coli SmpB. The tail-truncated diatom SmpB, the hypothetical product of a misspliced mRNA, was inactive in vivo. Tail-truncated E. coli SmpB was likewise inactive for tmRNA translation but was still able to bind ribosomes, and its affinity for tmRNA was only slightly diminished. This work suggests that SmpB is a universal cofactor of tmRNA. It also reveals a tail-dependent role for SmpB in tmRNA translation that supersedes a simple role of linking tmRNA to the ribosome, which the SmpB body alone could provide.
Jacob
Yannick
Y
Department of Biology, Indiana University, 1001 E. 3rd St., Bloomington, IN 47405, USA.
Sharkady
Stephen M
SM
Bhardwaj
Kanchan
K
Sanda
Alina
A
Williams
Kelly P
KP
D000483:Alleles; D000595:Amino Acid Sequence; D049148:Aminoacylation; D002467:Cell Nucleus / Q000378:metabolism*; D017377:Diatoms / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005057:Eukaryotic Cells / Q000378:metabolism; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000461:Rhodophyta / Q000235:genetics; D012270:Ribosomes / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics
D012329:RNA, Bacterial; D016601:RNA-Binding Proteins; C430328:small protein B; C096222:tmRNA