Biochemical analysis of human tRNAHis guanylyltransferase in mitochondrial tRNAHis maturation

Nakamura, Akiyoshi; Wang, Daole; Komatsu, Yasuo

doi:10.1016/j.bbrc.2018.07.150

Cited by 5 publications

(12 citation statements)

References 27 publications

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“…In almost all species, a single tRNA, tRNA His , is also modified at the 5′ end with an additional guanosine, termed G −1 [183], which acts as a recognition determinant for its cognate histidyl-tRNA synthetase [184]. Although the presence of G −1 has been shown to result from an altered RNase P cleavage of an encoded guanosine in bacteria [185] and yeast mitochondria [186], G −1 is not encoded in human mtDNA and its post-transcriptional addition on human mt-tRNAHis by THG1L has been demonstrated in vitro [187]. …”

Section: Mitochondrial Trna Modificationsmentioning

confidence: 99%

The mammalian mitochondrial epitranscriptome

Rebelo-Guiomar

Powell

Haute

et al. 2019

Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanism

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Correct expression of the mitochondrially-encoded genes is critical for the production of the components of the oxidative phosphorylation machinery. Post-transcriptional modifications of mitochondrial transcripts have been emerging as an important regulatory feature of mitochondrial gene expression. Here we review the current knowledge on how the mammalian mitochondrial epitranscriptome participates in regulating mitochondrial homeostasis. In particular, we focus on the latest breakthroughs made towards understanding the roles of the modified nucleotides in mitochondrially-encoded ribosomal and transfer RNAs, the enzymes responsible for introducing these modifications and on recent transcriptome-wide studies reporting modifications to mitochondrial messenger RNAs. This article is part of a Special Issue entitled: mRNA modifications in gene expression control edited by Dr. Matthias Soller and Dr. Rupert Fray.

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Section: Mitochondrial Trna Modificationsmentioning

confidence: 99%

The mammalian mitochondrial epitranscriptome

Rebelo-Guiomar

Powell

Haute

et al. 2019

Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanism

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“…In the current project we examined whether 5′RF/3′RF could be a substrate for HsThg1 as well as CaThg1. Cytoplasmic and mitochondrial tRNAs have A and C at the Y 73 opposite to G −1 ( Nakamura et al 2018a ). HsThg1 appears to be involved in the maturation of both tRNAs.…”

Section: Resultsmentioning

confidence: 99%

“…HsThg1 catalyzes the addition of G −1 to both human cytoplasmic (hc) and mitochondrial (hm) tRNA His . Human cytoplasmic tRNA His contains A at the Y 73 position, while mitochondrial tRNA His has C at this position ( Nakamura et al 2018a ). CaThg1 is involved in the maturation of cytoplasmic tRNA His with an A 73 but does not act on mitochondrial tRNA His because the G −1 of mitochondrial tRNA His in C. albicans is encoded in the mitochondrial genome the same as bacterial type tRNA His .…”

Section: Resultsmentioning

confidence: 99%

“…The amino-terminally His 6 -tagged HsThg1 without a mitochondrial targeting sequence and CaThg1 were overexpressed in E. coli BL21 (DE3)-pRARE2 (Novagen), and purified as previously described ( Nakamura et al 2013 , 2018a ). Purified Thg1 enzymes were dialyzed against 25 mM HEPES-Na pH 7.5, 500 mM NaCl, 4 mM MgCl 2 , 1 mM DTT, 50% (v/v) glycerol.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, it has been demonstrated that human Thg1 (HsThg1) catalyzes G −1 addition reactions in both human cytoplasmic (hc) and mitochondrial (hm) tRNA His . This observation suggests that HsThg1 could participate in both cytoplasmic and mitochondrial tRNA His maturation ( Nakamura et al 2018a ). In the cytoplasm of eukaryotes, Thg1 adds G −1 to the 5′-end of hctRNA His , opposite a conserved A 73 , resulting in a G:A 3′–5′ mismatch addition reaction.…”

Section: Introductionmentioning

confidence: 98%

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Analysis of GTP addition in the reverse (3′–5′) direction by human tRNA^His guanylyltransferase

Nakamura

Wang

Komatsu

2021

RNA

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Human tRNAHis guanylyltransferase (HsThg1) catalyzes the 3′–5′ addition of guanosine triphosphate (GTP) to the 5′-end (−1 position) of tRNAHis, producing mature tRNAHis. In human cells, cytoplasmic and mitochondrial tRNAHis have adenine (A) or cytidine (C), respectively, opposite to G−1. Little attention has been paid to the structural requirements of incoming GTP in 3′–5′ nucleotidyl addition by HsThg1. In this study, we evaluated the incorporation efficiencies of various GTP analogs by HsThg1 and compared the reaction mechanism with that of Candida albicans Thg1 (CaThg1). HsThg1 incorporated GTP opposite A or C in the template most efficiently. In contrast to CaThg1, HsThg1 could incorporate UTP opposite A, and guanosine diphosphate (GDP) opposite C. These results suggest that HsThg1 could transfer not only GTP, but also other NTPs, by forming Watson–Crick (WC) hydrogen bonds between the incoming NTP and the template base. On the basis of the molecular mechanism, HsThg1 succeeded in labeling the 5′-end of tRNAHis with biotinylated GTP. Structural analysis of HsThg1 was also performed in the presence of the mitochondrial tRNAHis. Structural comparison of HsThg1 with other Thg1 family enzymes suggested that the structural diversity of the carboxy-terminal domain of the Thg1 enzymes might be involved in the formation of WC base-pairing between the incoming GTP and template base. These findings provide new insights into an unidentified biological function of HsThg1 and also into the applicability of HsThg1 to the 5′-terminal modification of RNAs.

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Human Thg1 displays tRNA-inducible GTPase activity

Antika

Nazilah

Lee

et al. 2022

Nucleic Acids Research

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tRNAHis guanylyltransferase (Thg1) catalyzes the 3′-5′ incorporation of guanosine into position -1 (G-1) of tRNAHis. G-1 is unique to tRNAHis and is crucial for recognition by histidyl-tRNA synthetase (HisRS). Yeast Thg1 requires ATP for G-1 addition to tRNAHis opposite A73, whereas archaeal Thg1 requires either ATP or GTP for G-1 addition to tRNAHis opposite C73. Paradoxically, human Thg1 (HsThg1) can add G-1 to tRNAsHis with A73 (cytoplasmic) and C73 (mitochondrial). As N73 is immediately followed by a CCA end (positions 74–76), how HsThg1 prevents successive 3′-5′ incorporation of G-1/G-2/G-3 into mitochondrial tRNAHis (tRNAmHis) through a template-dependent mechanism remains a puzzle. We showed herein that mature native human tRNAmHis indeed contains only G-1. ATP was absolutely required for G-1 addition to tRNAmHis by HsThg1. Although HsThg1 could incorporate more than one GTP into tRNAmHisin vitro, a single-GTP incorporation prevailed when the relative GTP level was low. Surprisingly, HsThg1 possessed a tRNA-inducible GTPase activity, which could be inhibited by ATP. Similar activity was found in other high-eukaryotic dual-functional Thg1 enzymes, but not in yeast Thg1. This study suggests that HsThg1 may downregulate the level of GTP through its GTPase activity to prevent multiple-GTP incorporation into tRNAmHis.

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Biochemical analysis of human tRNAHis guanylyltransferase in mitochondrial tRNAHis maturation

Cited by 5 publications

References 27 publications

The mammalian mitochondrial epitranscriptome

The mammalian mitochondrial epitranscriptome

Analysis of GTP addition in the reverse (3′–5′) direction by human tRNA^His guanylyltransferase

Human Thg1 displays tRNA-inducible GTPase activity

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Biochemical analysis of human tRNAHis guanylyltransferase in mitochondrial tRNAHis maturation

Cited by 5 publications

References 27 publications

The mammalian mitochondrial epitranscriptome

The mammalian mitochondrial epitranscriptome

Analysis of GTP addition in the reverse (3′–5′) direction by human tRNAHis guanylyltransferase

Human Thg1 displays tRNA-inducible GTPase activity

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Analysis of GTP addition in the reverse (3′–5′) direction by human tRNA^His guanylyltransferase