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Research article

The fission yeast Schizosaccharomyces pombe has two distinct tRNase ZLs encoded by two different genes and differentially targeted to the nucleus and mitochondria

Xuhua Gan, Jing Yang, Jun Li, Haiyan Yu, Hongmei Dai, Jinyu Liu, Ying Huang
Biochemical Journal Apr 01, 2011, 435 (1) 103-111; DOI: 10.1042/BJ20101619
Xuhua Gan
Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, School of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China
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Jing Yang
Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, School of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China
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Jun Li
Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, School of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China
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Haiyan Yu
Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, School of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China
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Hongmei Dai
Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, School of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China
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Jinyu Liu
Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, School of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China
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Ying Huang
Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, School of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China
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  • For correspondence: yhuang@njnu.edu.cn
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Abstract

tRNase Z is the endonuclease that is involved in tRNA 3′-end maturation by removal of the 3′-trailer sequences from tRNA precursors. Most eukaryotes examined to date, including the budding yeast Saccharomyces cerevisiae and humans, have a single long form of tRNase Z (tRNase ZL). In contrast, the fission yeast Schizosaccharomyces pombe contains two candidate tRNase ZLs encoded by the essential genes sptrz1+ and sptrz2+. In the present study, we have expressed recombinant SpTrz1p and SpTrz2p in S. pombe. Both recombinant proteins possess precursor tRNA 3′-endonucleolytic activity in vitro. SpTrz1p localizes to the nucleus and has a simian virus 40 NLS (nuclear localization signal)-like NLS at its N-terminus, which contains four consecutive arginine and lysine residues between residues 208 and 211 that are critical for the NLS function. In contrast, SpTrz2p is a mitochondrial protein with an N-terminal MTS (mitochondrial-targeting signal). High-level overexpression of sptrz1+ has no detectable phenotypes. In contrast, strong overexpression of sptrz2+ is lethal in wild-type cells and results in morphological abnormalities, including swollen and round cells, demonstrating that the correct expression level of sptrz2+ is critical. The present study provides evidence for partitioning of tRNase Z function between two different proteins in S. pombe, although we cannot rule out specialized functions for each protein.

  • endonuclease
  • 3′-end processing
  • post-transcriptional processing
  • tRNA precursor (pre-tRNA)
  • tRNase Z

Abbreviations: aa, amino acids; DAPI, 4′,6-diamidino-2-phenylindole; DIC, differential interference contrast; DTT, dithiothreitol; EGFP, enhanced green fluorescent protein; EMM, Edinburgh minimal medium; GST, glutathione transferase; MBL, metallo-β-lactamase; miRNA, microRNA; MTS, mitochondrial-targeting signal; NLS, nuclear localization signal; nmt1, no message in thiamine; pre-tRNA, tRNA precursor; ScTrz1p, Saccharomyces cerevisiae tRNase ZL; SpTrz1p, S. pombe tRNase ZL; SV40, simian virus 40; tRNase, ZL, long form of tRNase Z; tRNase, ZS, short form of tRNase Z; YES, yeast extract medium plus supplements

  • © The Authors Journal compilation © 2011 Biochemical Society
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April 2011

Volume: 435 Issue: 1

Biochemical Journal: 435 (1)
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The fission yeast Schizosaccharomyces pombe has two distinct tRNase ZLs encoded by two different genes and differentially targeted to the nucleus and mitochondria
Xuhua Gan, Jing Yang, Jun Li, Haiyan Yu, Hongmei Dai, Jinyu Liu, Ying Huang
Biochemical Journal Apr 2011, 435 (1) 103-111; DOI: 10.1042/BJ20101619
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The fission yeast Schizosaccharomyces pombe has two distinct tRNase ZLs encoded by two different genes and differentially targeted to the nucleus and mitochondria
Xuhua Gan, Jing Yang, Jun Li, Haiyan Yu, Hongmei Dai, Jinyu Liu, Ying Huang
Biochemical Journal Apr 2011, 435 (1) 103-111; DOI: 10.1042/BJ20101619

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Keywords

endonuclease
3′-end processing
post-transcriptional processing
tRNA precursor (pre-tRNA)
tRNase Z

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