Crystal structure of tRNAHis guanylyltransferase from Saccharomyces cerevisiae

Kitaik Lee; Eun Hye Lee; Jonghyeon Son; Kwang Yeon Hwang

doi:10.1016/j.bbrc.2017.06.054

Crystal structure of tRNA^His guanylyltransferase from Saccharomyces cerevisiae

Kitaik Lee, Eun Hye Lee, Jonghyeon Son, Kwang Yeon Hwang

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

Abstract

tRNA maturation involves several steps, including processing, splicing, CCA addition, and posttranscriptional modifications. tRNA^His guanylyltransferase (Thg1) is the only enzyme known to catalyze templated nucleotide addition in the 3′–5′ direction, unlike other DNA and RNA polymerases. For a better understanding of its unique catalytic mechanism at the molecular level, we determined the crystal structure of GTP-bound Thg1 from Saccharomyces cerevisiae at the maximum resolution of 3.0 Å. The structure revealed the enzyme to have a tetrameric conformation that is well conserved among different species, and the GTP molecule was clearly bound at the active site, coordinating with two magnesium ions. In addition, two flexible protomers at the potential binding site (PBS) for tRNA^His were observed. We suggest that the PBS of the tetramer could also be one of the sites for interaction with partner proteins.

Original language	English
Pages (from-to)	400-405
Number of pages	6
Journal	Biochemical and biophysical research communications
Volume	490
Issue number	2
DOIs	https://doi.org/10.1016/j.bbrc.2017.06.054
Publication status	Published - 2017 Aug 19

Bibliographical note

Funding Information:
We thank the supporting staff of beamline NW12A of the Photon Factory (Tsukuba, Japan) and beamline 5C-SBII of Pohang Accelerator Light Source (Pohang, Korea) for their help with the data collection. This work was supported by grants from the National Research Foundation of Korea (2017R1A2B2005666). KYH was supported by Korea University grants.

Publisher Copyright:
© 2017 Elsevier Inc.

Keywords

GTP
Posttranscriptional modifications
Reverse polymerization
Thg1
X-ray structure

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.bbrc.2017.06.054

Cite this

@article{3eed55cf69de4f90b89295d890e78c6f,

title = "Crystal structure of tRNAHis guanylyltransferase from Saccharomyces cerevisiae",

abstract = "tRNA maturation involves several steps, including processing, splicing, CCA addition, and posttranscriptional modifications. tRNAHis guanylyltransferase (Thg1) is the only enzyme known to catalyze templated nucleotide addition in the 3′–5′ direction, unlike other DNA and RNA polymerases. For a better understanding of its unique catalytic mechanism at the molecular level, we determined the crystal structure of GTP-bound Thg1 from Saccharomyces cerevisiae at the maximum resolution of 3.0 {\AA}. The structure revealed the enzyme to have a tetrameric conformation that is well conserved among different species, and the GTP molecule was clearly bound at the active site, coordinating with two magnesium ions. In addition, two flexible protomers at the potential binding site (PBS) for tRNAHis were observed. We suggest that the PBS of the tetramer could also be one of the sites for interaction with partner proteins.",

keywords = "GTP, Posttranscriptional modifications, Reverse polymerization, Thg1, X-ray structure",

author = "Kitaik Lee and Lee, {Eun Hye} and Jonghyeon Son and Hwang, {Kwang Yeon}",

note = "Funding Information: We thank the supporting staff of beamline NW12A of the Photon Factory (Tsukuba, Japan) and beamline 5C-SBII of Pohang Accelerator Light Source (Pohang, Korea) for their help with the data collection. This work was supported by grants from the National Research Foundation of Korea (2017R1A2B2005666). KYH was supported by Korea University grants. Publisher Copyright: {\textcopyright} 2017 Elsevier Inc.",

year = "2017",

month = aug,

day = "19",

doi = "10.1016/j.bbrc.2017.06.054",

language = "English",

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AU - Lee, Kitaik

AU - Lee, Eun Hye

AU - Son, Jonghyeon

AU - Hwang, Kwang Yeon

N1 - Funding Information: We thank the supporting staff of beamline NW12A of the Photon Factory (Tsukuba, Japan) and beamline 5C-SBII of Pohang Accelerator Light Source (Pohang, Korea) for their help with the data collection. This work was supported by grants from the National Research Foundation of Korea (2017R1A2B2005666). KYH was supported by Korea University grants. Publisher Copyright: © 2017 Elsevier Inc.

PY - 2017/8/19

Y1 - 2017/8/19

N2 - tRNA maturation involves several steps, including processing, splicing, CCA addition, and posttranscriptional modifications. tRNAHis guanylyltransferase (Thg1) is the only enzyme known to catalyze templated nucleotide addition in the 3′–5′ direction, unlike other DNA and RNA polymerases. For a better understanding of its unique catalytic mechanism at the molecular level, we determined the crystal structure of GTP-bound Thg1 from Saccharomyces cerevisiae at the maximum resolution of 3.0 Å. The structure revealed the enzyme to have a tetrameric conformation that is well conserved among different species, and the GTP molecule was clearly bound at the active site, coordinating with two magnesium ions. In addition, two flexible protomers at the potential binding site (PBS) for tRNAHis were observed. We suggest that the PBS of the tetramer could also be one of the sites for interaction with partner proteins.

AB - tRNA maturation involves several steps, including processing, splicing, CCA addition, and posttranscriptional modifications. tRNAHis guanylyltransferase (Thg1) is the only enzyme known to catalyze templated nucleotide addition in the 3′–5′ direction, unlike other DNA and RNA polymerases. For a better understanding of its unique catalytic mechanism at the molecular level, we determined the crystal structure of GTP-bound Thg1 from Saccharomyces cerevisiae at the maximum resolution of 3.0 Å. The structure revealed the enzyme to have a tetrameric conformation that is well conserved among different species, and the GTP molecule was clearly bound at the active site, coordinating with two magnesium ions. In addition, two flexible protomers at the potential binding site (PBS) for tRNAHis were observed. We suggest that the PBS of the tetramer could also be one of the sites for interaction with partner proteins.

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KW - X-ray structure

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