Trans-editing of Cys-tRNAPro by Haemophilus influenzae YbaK protein.
Prolyl-tRNA synthetases (ProRSs) from all three domains of life have been shown to misactivate cysteine and to mischarge cysteine onto tRNAPro. Although most bacterial ProRSs possess an amino acid editing domain that deacylates mischarged Ala-tRNAPro, editing of Cys-tRNAPro has not been demonstrated and a double-sieve mechanism of editing does not appear to be sufficient to eliminate all misacylated tRNAPro species from the cell. It was recently shown that a ProRS paralog, the YbaK protein from Haemophilus influenzae, which is homologous to the ProRS editing domain, is capable of weakly deacylating Ala-tRNAPro. This function appears to be redundant with that of its corresponding ProRS, which contains a canonical bacterial editing domain. In the present study, we test the specificity of editing by H. influenzae YbaK and show that it efficiently edits Cys-tRNAPro and that a conserved Lys residue is essential for this activity. These findings represent the first example of an editing domain paralog possessing altered specificity and suggest that similar autonomous editing domains could act upon different mischarged tRNAs thus providing cells with enhanced proofreading potential. This work also suggests a novel mechanism of editing wherein a third sieve is used to clear Cys-tRNAPro in at least some organisms.
An
Songon
S
Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Musier-Forsyth
Karin
K
D000604:Amino Acyl-tRNA Synthetases / Q000502:physiology; D001426:Bacterial Proteins / Q000502:physiology*; D003545:Cysteine / Q000737:chemistry; D003553:Cystine / Q000737:chemistry; D006193:Haemophilus influenzae / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry; D017434:Protein Structure, Tertiary; D017393:RNA Editing; D012343:RNA, Transfer / Q000737:chemistry; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry*; D012361:RNA, Transfer, Pro / Q000737:chemistry*; D013997:Time Factors
D001426:Bacterial Proteins; D012346:RNA, Transfer, Amino Acyl; D012361:RNA, Transfer, Pro; C481278:YbaK protein, Haemophilus influenzae; D003553:Cystine; D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases; C100134:prolyl T RNA synthetase; D008239:Lysine; D003545:Cysteine