Differential modes of transfer RNASer recognition in Methanosarcina barkeri.
Two dissimilar seryl-transfer RNA (tRNA) synthetases (SerRSs) exist in Methanosarcina barkeri, one of bacterial type and the other resembling SerRSs present only in some methanogenic archaea. To investigate the requirements of these enzymes for tRNASer recognition, serylation of variant transcripts of M. barkeri tRNASer was kinetically analyzed in vitro with pure enzyme preparations. Characteristically for the serine system, the length of the variable arm was shown to be crucial for both enzymes, as was the identity of the discriminator base (G73). Moreover, a novel determinant for the specific tRNASer recognition was identified as the anticodon stem base pair G30:C40; its contribution to the efficiency of serylation was remarkable for both SerRSs. However, despite these similarities, the two SerRSs do not possess a uniform mode of tRNASer recognition, and additional determinants are necessary for serylation specificity by the methanogenic enzyme. In particular, the methanogenic SerRS relies on G1:C72 identity and on the number of unpaired nucleotides at the base of the variable stem for tRNASer recognition, unlike its bacterial type counterpart. We propose that such a distinction between the two enzymes in tRNASer identity determinants reflects their evolutionary pathways, hence attesting to their diversity.
Korencic
Dragana
D
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520-8114, USA.
Polycarpo
Carla
C
Weygand-Durasevic
Ivana
I
Söll
Dieter
D
D001483:Base Sequence; D003001:Cloning, Molecular; D019143:Evolution, Molecular; D007700:Kinetics; D017021:Methanosarcina barkeri / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D012343:RNA, Transfer / Q000737:chemistry; D012362:RNA, Transfer, Ser / Q000378:metabolism*; D012694:Serine / Q000737:chemistry; D012714:Serine-tRNA Ligase / Q000737:chemistry* / Q000235:genetics*
D012362:RNA, Transfer, Ser; D012694:Serine; D012343:RNA, Transfer; D012714:Serine-tRNA Ligase