A Temporal Order in 5′- and 3′- Processing of Eukaryotic tRNAHis
For flawless translation of mRNA sequence into protein, tRNAs must undergo a series of essential maturation steps to be properly recognized and aminoacylated by aminoacyl-tRNA synthetase, and subsequently utilized by the ribosome. While all tRNAs carry a 3′-terminal CCA sequence that inclu...
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doaj-4c07e09ffbf64ce48befb4e3479b06152020-11-25T00:55:11ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-03-01206138410.3390/ijms20061384ijms20061384A Temporal Order in 5′- and 3′- Processing of Eukaryotic tRNAHisMarie-Theres Pöhler0Tracy M. Roach1Heike Betat2Jane E. Jackman3Mario Mörl4Institute for Biochemistry, Leipzig University, Brüderstraße 34, 04103 Leipzig, GermanyCenter for RNA Biology and Ohio State Biochemistry Program, Department of Chemistry and Biochemistry, the Ohio State University, Columbus, OH 43210, USAInstitute for Biochemistry, Leipzig University, Brüderstraße 34, 04103 Leipzig, GermanyCenter for RNA Biology and Ohio State Biochemistry Program, Department of Chemistry and Biochemistry, the Ohio State University, Columbus, OH 43210, USAInstitute for Biochemistry, Leipzig University, Brüderstraße 34, 04103 Leipzig, GermanyFor flawless translation of mRNA sequence into protein, tRNAs must undergo a series of essential maturation steps to be properly recognized and aminoacylated by aminoacyl-tRNA synthetase, and subsequently utilized by the ribosome. While all tRNAs carry a 3′-terminal CCA sequence that includes the site of aminoacylation, the additional 5′-G-1 position is a unique feature of most histidine tRNA species, serving as an identity element for the corresponding synthetase. In eukaryotes including yeast, both 3′-CCA and 5′-G-1 are added post-transcriptionally by tRNA nucleotidyltransferase and tRNAHis guanylyltransferase, respectively. Hence, it is possible that these two cytosolic enzymes compete for the same tRNA. Here, we investigate substrate preferences associated with CCA and G-1-addition to yeast cytosolic tRNAHis, which might result in a temporal order to these important processing events. We show that tRNA nucleotidyltransferase accepts tRNAHis transcripts independent of the presence of G-1; however, tRNAHis guanylyltransferase clearly prefers a substrate carrying a CCA terminus. Although many tRNA maturation steps can occur in a rather random order, our data demonstrate a likely pathway where CCA-addition precedes G-1 incorporation in S. cerevisiae. Evidently, the 3′-CCA triplet and a discriminator position A73 act as positive elements for G-1 incorporation, ensuring the fidelity of G-1 addition.http://www.mdpi.com/1422-0067/20/6/1384CCA-additionG-1 residuetRNAHis guanylyltransferasetRNAHistRNA maturationtRNA nucleotidyltransferase |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Marie-Theres Pöhler Tracy M. Roach Heike Betat Jane E. Jackman Mario Mörl |
spellingShingle |
Marie-Theres Pöhler Tracy M. Roach Heike Betat Jane E. Jackman Mario Mörl A Temporal Order in 5′- and 3′- Processing of Eukaryotic tRNAHis International Journal of Molecular Sciences CCA-addition G-1 residue tRNAHis guanylyltransferase tRNAHis tRNA maturation tRNA nucleotidyltransferase |
author_facet |
Marie-Theres Pöhler Tracy M. Roach Heike Betat Jane E. Jackman Mario Mörl |
author_sort |
Marie-Theres Pöhler |
title |
A Temporal Order in 5′- and 3′- Processing of Eukaryotic tRNAHis |
title_short |
A Temporal Order in 5′- and 3′- Processing of Eukaryotic tRNAHis |
title_full |
A Temporal Order in 5′- and 3′- Processing of Eukaryotic tRNAHis |
title_fullStr |
A Temporal Order in 5′- and 3′- Processing of Eukaryotic tRNAHis |
title_full_unstemmed |
A Temporal Order in 5′- and 3′- Processing of Eukaryotic tRNAHis |
title_sort |
temporal order in 5′- and 3′- processing of eukaryotic trnahis |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2019-03-01 |
description |
For flawless translation of mRNA sequence into protein, tRNAs must undergo a series of essential maturation steps to be properly recognized and aminoacylated by aminoacyl-tRNA synthetase, and subsequently utilized by the ribosome. While all tRNAs carry a 3′-terminal CCA sequence that includes the site of aminoacylation, the additional 5′-G-1 position is a unique feature of most histidine tRNA species, serving as an identity element for the corresponding synthetase. In eukaryotes including yeast, both 3′-CCA and 5′-G-1 are added post-transcriptionally by tRNA nucleotidyltransferase and tRNAHis guanylyltransferase, respectively. Hence, it is possible that these two cytosolic enzymes compete for the same tRNA. Here, we investigate substrate preferences associated with CCA and G-1-addition to yeast cytosolic tRNAHis, which might result in a temporal order to these important processing events. We show that tRNA nucleotidyltransferase accepts tRNAHis transcripts independent of the presence of G-1; however, tRNAHis guanylyltransferase clearly prefers a substrate carrying a CCA terminus. Although many tRNA maturation steps can occur in a rather random order, our data demonstrate a likely pathway where CCA-addition precedes G-1 incorporation in S. cerevisiae. Evidently, the 3′-CCA triplet and a discriminator position A73 act as positive elements for G-1 incorporation, ensuring the fidelity of G-1 addition. |
topic |
CCA-addition G-1 residue tRNAHis guanylyltransferase tRNAHis tRNA maturation tRNA nucleotidyltransferase |
url |
http://www.mdpi.com/1422-0067/20/6/1384 |
work_keys_str_mv |
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1725231585814380544 |