Trehalose 6-phosphate synthase and trehalose 6-phosphate phosphatase from Nicotiana tabacum function in trehalose biosynthesis and environmental stress tolerance of yeast

Takeshi Machida, Ken-ichi Honjoh, A. S O Ayuko, Maiko Yamamoto, H. O. Masayoshi, Takahisa Miyamoto

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

To investigate functions of trehalose 6-phosphate synthase (TPS) and trehalose 6-phosphate phosphatase (TPP) from a tobacco plant, Nicotiana tabacum, the corresponding cDNA clones were isolated. Those genes were designated NtTPS and NtTPP, respectively. NtTPS included a N-terminal extension according to comparison of deduced amino acid sequence of NtTPS with those of TPSs from Escherichia coli and yeast. The NtTPS was genetically modified to lack a region for the N-terminal extension and the modified gene was designated ΔNNtTPS. The genes were expressed in yeast tps1 mutant as two separate proteins and as a NtTPS (or ΔNNtTPS)-NtTPP fusion protein. Western blot analysis showed that the NtTPS, NtTPP, and NtTPS-NtTPP were expressed abundantly in yeast, while the ANNtTPS and ANNtTPSNtTPP were not detected. Interestingly, high levels of trehalose were accumulated in yeast expressing ΔNNtTPS and ΔNNtTPS-NtTPP in spite of their low-level expressions. Furthermore, stress tolerances of yeast against osmotic, freezing-thawing, and heat stresses were significantly improved by the expression of the tobacco gene, and the increased levels in tolerance were proportional to their trehalose levels. Our results showed that NtTPS and NtTPP functioned in trehalose synthesis by the removal of N-terminal extension of NtTPS and several environmental stress tolerances.

Original languageEnglish
Pages (from-to)261-268
Number of pages8
JournalJournal of the Faculty of Agriculture, Kyushu University
Volume55
Issue number2
Publication statusPublished - Oct 1 2010

Fingerprint

Trehalose
trehalose
stress tolerance
Nicotiana tabacum
Tobacco
Yeasts
biosynthesis
yeasts
tobacco
genes
Genes
freeze-thaw cycles
Freezing
heat stress
Amino Acid Sequence
Western blotting
Proteins
amino acid sequences
proteins
Complementary DNA

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Agronomy and Crop Science

Cite this

@article{e049ac227dbb4e1d8151f6345c487523,
title = "Trehalose 6-phosphate synthase and trehalose 6-phosphate phosphatase from Nicotiana tabacum function in trehalose biosynthesis and environmental stress tolerance of yeast",
abstract = "To investigate functions of trehalose 6-phosphate synthase (TPS) and trehalose 6-phosphate phosphatase (TPP) from a tobacco plant, Nicotiana tabacum, the corresponding cDNA clones were isolated. Those genes were designated NtTPS and NtTPP, respectively. NtTPS included a N-terminal extension according to comparison of deduced amino acid sequence of NtTPS with those of TPSs from Escherichia coli and yeast. The NtTPS was genetically modified to lack a region for the N-terminal extension and the modified gene was designated ΔNNtTPS. The genes were expressed in yeast tps1 mutant as two separate proteins and as a NtTPS (or ΔNNtTPS)-NtTPP fusion protein. Western blot analysis showed that the NtTPS, NtTPP, and NtTPS-NtTPP were expressed abundantly in yeast, while the ANNtTPS and ANNtTPSNtTPP were not detected. Interestingly, high levels of trehalose were accumulated in yeast expressing ΔNNtTPS and ΔNNtTPS-NtTPP in spite of their low-level expressions. Furthermore, stress tolerances of yeast against osmotic, freezing-thawing, and heat stresses were significantly improved by the expression of the tobacco gene, and the increased levels in tolerance were proportional to their trehalose levels. Our results showed that NtTPS and NtTPP functioned in trehalose synthesis by the removal of N-terminal extension of NtTPS and several environmental stress tolerances.",
author = "Takeshi Machida and Ken-ichi Honjoh and Ayuko, {A. S O} and Maiko Yamamoto and Masayoshi, {H. O.} and Takahisa Miyamoto",
year = "2010",
month = "10",
day = "1",
language = "English",
volume = "55",
pages = "261--268",
journal = "Journal of the Faculty of Agriculture, Kyushu University",
issn = "0023-6152",
publisher = "Faculty of Agriculture, Kyushu University",
number = "2",

}

TY - JOUR

T1 - Trehalose 6-phosphate synthase and trehalose 6-phosphate phosphatase from Nicotiana tabacum function in trehalose biosynthesis and environmental stress tolerance of yeast

AU - Machida, Takeshi

AU - Honjoh, Ken-ichi

AU - Ayuko, A. S O

AU - Yamamoto, Maiko

AU - Masayoshi, H. O.

AU - Miyamoto, Takahisa

PY - 2010/10/1

Y1 - 2010/10/1

N2 - To investigate functions of trehalose 6-phosphate synthase (TPS) and trehalose 6-phosphate phosphatase (TPP) from a tobacco plant, Nicotiana tabacum, the corresponding cDNA clones were isolated. Those genes were designated NtTPS and NtTPP, respectively. NtTPS included a N-terminal extension according to comparison of deduced amino acid sequence of NtTPS with those of TPSs from Escherichia coli and yeast. The NtTPS was genetically modified to lack a region for the N-terminal extension and the modified gene was designated ΔNNtTPS. The genes were expressed in yeast tps1 mutant as two separate proteins and as a NtTPS (or ΔNNtTPS)-NtTPP fusion protein. Western blot analysis showed that the NtTPS, NtTPP, and NtTPS-NtTPP were expressed abundantly in yeast, while the ANNtTPS and ANNtTPSNtTPP were not detected. Interestingly, high levels of trehalose were accumulated in yeast expressing ΔNNtTPS and ΔNNtTPS-NtTPP in spite of their low-level expressions. Furthermore, stress tolerances of yeast against osmotic, freezing-thawing, and heat stresses were significantly improved by the expression of the tobacco gene, and the increased levels in tolerance were proportional to their trehalose levels. Our results showed that NtTPS and NtTPP functioned in trehalose synthesis by the removal of N-terminal extension of NtTPS and several environmental stress tolerances.

AB - To investigate functions of trehalose 6-phosphate synthase (TPS) and trehalose 6-phosphate phosphatase (TPP) from a tobacco plant, Nicotiana tabacum, the corresponding cDNA clones were isolated. Those genes were designated NtTPS and NtTPP, respectively. NtTPS included a N-terminal extension according to comparison of deduced amino acid sequence of NtTPS with those of TPSs from Escherichia coli and yeast. The NtTPS was genetically modified to lack a region for the N-terminal extension and the modified gene was designated ΔNNtTPS. The genes were expressed in yeast tps1 mutant as two separate proteins and as a NtTPS (or ΔNNtTPS)-NtTPP fusion protein. Western blot analysis showed that the NtTPS, NtTPP, and NtTPS-NtTPP were expressed abundantly in yeast, while the ANNtTPS and ANNtTPSNtTPP were not detected. Interestingly, high levels of trehalose were accumulated in yeast expressing ΔNNtTPS and ΔNNtTPS-NtTPP in spite of their low-level expressions. Furthermore, stress tolerances of yeast against osmotic, freezing-thawing, and heat stresses were significantly improved by the expression of the tobacco gene, and the increased levels in tolerance were proportional to their trehalose levels. Our results showed that NtTPS and NtTPP functioned in trehalose synthesis by the removal of N-terminal extension of NtTPS and several environmental stress tolerances.

UR - http://www.scopus.com/inward/record.url?scp=78549274719&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78549274719&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:78549274719

VL - 55

SP - 261

EP - 268

JO - Journal of the Faculty of Agriculture, Kyushu University

JF - Journal of the Faculty of Agriculture, Kyushu University

SN - 0023-6152

IS - 2

ER -