Template-dependent 3'-5' nucleotide addition is a shared feature of tRNAHis guanylyltransferase enzymes from multiple domains of life.
The presence of an additional 5' guanosine residue (G(-1)) is a unique feature of tRNA(His). G(-1) is incorporated posttranscriptionally in eukarya via an unusual 3'-5' nucleotide addition reaction catalyzed by the tRNA(His) guanylyltransferase (Thg1). Yeast Thg1 catalyzes an unexpected second activity: Watson-Crick-dependent 3'-5' nucleotide addition that occurs in the opposite direction to nucleotide addition by all known DNA and RNA polymerases. This discovery led to the hypothesis that there are alternative roles for Thg1 family members that take advantage of this unusual enzymatic activity. Here we show that archaeal homologs of Thg1 catalyze G(-1) addition, in vitro and in vivo in yeast, but only in a templated reaction, i.e. with tRNA(His) substrates that contain a C(73) discriminator nucleotide. Because tRNA(His) from archaea contains C(73), these findings are consistent with a physiological function for templated nucleotide addition in archaeal tRNA(His) maturation. Moreover, unlike yeast Thg1, archaeal Thg1 enzymes also exhibit a preference for template-dependent U(-1) addition to A(73)-containing tRNA(His). Taken together, these results demonstrate that Watson-Crick template-dependent 3'-5' nucleotide addition is a shared catalytic activity exhibited by Thg1 family members from multiple domains of life, and therefore, that this unusual reaction may constitute an ancestral activity present in the earliest members of the Thg1 enzyme family.
Abad
Maria G
MG
Department of Biochemistry and Center for RNA Biology, The Ohio State University, Columbus, OH 43210, USA.
Rao
Bhalchandra S
BS
Jackman
Jane E
JE
D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009690:Nucleic Acid Conformation; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D012354:RNA, Transfer, His / Q000737:chemistry / Q000378:metabolism*; D015003:Yeasts / Q000201:enzymology
D019843:Archaeal Proteins; D001426:Bacterial Proteins; D005656:Fungal Proteins; D012354:RNA, Transfer, His; D009713:Nucleotidyltransferases; C020842:guanylyltransferase