Candida tropicalis expresses two mitochondrial 2-enoyl thioester reductases that are able to form both homodimers and heterodimers

Juha Torkko, Kari Koivuranta, Alexander Kastaniotis, Tomi Airenne, Tuomo Glumoff, Mika Ilves, Andreas Hartig, Aner Gurvitz, Kalervo Hiltunen (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

24 Citations (Scopus)

Abstract

Here we report on the cloning of a Candida tropicalis gene, ETR2, that is closely related to ETR1. Both genes encode enzymatically active 2-enoyl thioester reductases involved in mitochondrial synthesis of fatty acids (fatty acid synthesis type II) and respiratory competence. The 5′- and 3′-flanking (coding) regions of ETR2 and ETR1 are about 90% (97%) identical, indicating that the genes have evolved via gene duplication. The gene products differ in three amino acid residues: Ile67 (Val), Ala92 (Thr), and Lys251 (Arg) in Etr2p (Etr1p). Quantitative PCR analysis and reverse transcriptase-PCR indicated that both genes were expressed about equally in fermenting and ETR1 predominantly respiring yeast cells. Like the situation with ETR1, expression of ETR2 in respiration-deficient Saccharomyces cerevisiae mutant cells devoid of Ybr026p/Etr1p was able to restore growth on glycerol. Triclosan that is used as an antibacterial agent against fatty acid synthesis type II 2-enoyl thioester reductases inhibited growth of FabI overexpressing mutant yeast cells but was not able to inhibit respiratory growth of the ETR2- or ETR1-complemented mutant yeast cells. Resolving of crystal structures obtained via Etr2p and Etr1p co-crystallization indicated that all possible dimer variants occur in the same asymmetric unit, suggesting that similar dimer formation also takes place in vivo.
Original languageEnglish
Pages (from-to)41213-41220
Number of pages8
JournalJournal of Biological Chemistry
Volume278
Issue number42
DOIs
Publication statusPublished - 2003
MoE publication typeA1 Journal article-refereed

Fingerprint

Candida tropicalis
Candida
Genes
Yeast
Fatty Acids
Yeasts
Cells
Dimers
Growth
Triclosan
3' Flanking Region
Gene Duplication
Crystallization
Reverse Transcriptase Polymerase Chain Reaction
Cloning
RNA-Directed DNA Polymerase
Glycerol
Mental Competency
Saccharomyces cerevisiae
Organism Cloning

Keywords

  • Candida tropicalis
  • gene cloning
  • genes
  • reductase
  • fatty acids

Cite this

Torkko, Juha ; Koivuranta, Kari ; Kastaniotis, Alexander ; Airenne, Tomi ; Glumoff, Tuomo ; Ilves, Mika ; Hartig, Andreas ; Gurvitz, Aner ; Hiltunen, Kalervo. / Candida tropicalis expresses two mitochondrial 2-enoyl thioester reductases that are able to form both homodimers and heterodimers. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 42. pp. 41213-41220.
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title = "Candida tropicalis expresses two mitochondrial 2-enoyl thioester reductases that are able to form both homodimers and heterodimers",
abstract = "Here we report on the cloning of a Candida tropicalis gene, ETR2, that is closely related to ETR1. Both genes encode enzymatically active 2-enoyl thioester reductases involved in mitochondrial synthesis of fatty acids (fatty acid synthesis type II) and respiratory competence. The 5′- and 3′-flanking (coding) regions of ETR2 and ETR1 are about 90{\%} (97{\%}) identical, indicating that the genes have evolved via gene duplication. The gene products differ in three amino acid residues: Ile67 (Val), Ala92 (Thr), and Lys251 (Arg) in Etr2p (Etr1p). Quantitative PCR analysis and reverse transcriptase-PCR indicated that both genes were expressed about equally in fermenting and ETR1 predominantly respiring yeast cells. Like the situation with ETR1, expression of ETR2 in respiration-deficient Saccharomyces cerevisiae mutant cells devoid of Ybr026p/Etr1p was able to restore growth on glycerol. Triclosan that is used as an antibacterial agent against fatty acid synthesis type II 2-enoyl thioester reductases inhibited growth of FabI overexpressing mutant yeast cells but was not able to inhibit respiratory growth of the ETR2- or ETR1-complemented mutant yeast cells. Resolving of crystal structures obtained via Etr2p and Etr1p co-crystallization indicated that all possible dimer variants occur in the same asymmetric unit, suggesting that similar dimer formation also takes place in vivo.",
keywords = "Candida tropicalis, gene cloning, genes, reductase, fatty acids",
author = "Juha Torkko and Kari Koivuranta and Alexander Kastaniotis and Tomi Airenne and Tuomo Glumoff and Mika Ilves and Andreas Hartig and Aner Gurvitz and Kalervo Hiltunen",
year = "2003",
doi = "10.1074/jbc.M307664200",
language = "English",
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pages = "41213--41220",
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Torkko, J, Koivuranta, K, Kastaniotis, A, Airenne, T, Glumoff, T, Ilves, M, Hartig, A, Gurvitz, A & Hiltunen, K 2003, 'Candida tropicalis expresses two mitochondrial 2-enoyl thioester reductases that are able to form both homodimers and heterodimers', Journal of Biological Chemistry, vol. 278, no. 42, pp. 41213-41220. https://doi.org/10.1074/jbc.M307664200

Candida tropicalis expresses two mitochondrial 2-enoyl thioester reductases that are able to form both homodimers and heterodimers. / Torkko, Juha; Koivuranta, Kari; Kastaniotis, Alexander; Airenne, Tomi; Glumoff, Tuomo; Ilves, Mika; Hartig, Andreas; Gurvitz, Aner; Hiltunen, Kalervo (Corresponding Author).

In: Journal of Biological Chemistry, Vol. 278, No. 42, 2003, p. 41213-41220.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Candida tropicalis expresses two mitochondrial 2-enoyl thioester reductases that are able to form both homodimers and heterodimers

AU - Torkko, Juha

AU - Koivuranta, Kari

AU - Kastaniotis, Alexander

AU - Airenne, Tomi

AU - Glumoff, Tuomo

AU - Ilves, Mika

AU - Hartig, Andreas

AU - Gurvitz, Aner

AU - Hiltunen, Kalervo

PY - 2003

Y1 - 2003

N2 - Here we report on the cloning of a Candida tropicalis gene, ETR2, that is closely related to ETR1. Both genes encode enzymatically active 2-enoyl thioester reductases involved in mitochondrial synthesis of fatty acids (fatty acid synthesis type II) and respiratory competence. The 5′- and 3′-flanking (coding) regions of ETR2 and ETR1 are about 90% (97%) identical, indicating that the genes have evolved via gene duplication. The gene products differ in three amino acid residues: Ile67 (Val), Ala92 (Thr), and Lys251 (Arg) in Etr2p (Etr1p). Quantitative PCR analysis and reverse transcriptase-PCR indicated that both genes were expressed about equally in fermenting and ETR1 predominantly respiring yeast cells. Like the situation with ETR1, expression of ETR2 in respiration-deficient Saccharomyces cerevisiae mutant cells devoid of Ybr026p/Etr1p was able to restore growth on glycerol. Triclosan that is used as an antibacterial agent against fatty acid synthesis type II 2-enoyl thioester reductases inhibited growth of FabI overexpressing mutant yeast cells but was not able to inhibit respiratory growth of the ETR2- or ETR1-complemented mutant yeast cells. Resolving of crystal structures obtained via Etr2p and Etr1p co-crystallization indicated that all possible dimer variants occur in the same asymmetric unit, suggesting that similar dimer formation also takes place in vivo.

AB - Here we report on the cloning of a Candida tropicalis gene, ETR2, that is closely related to ETR1. Both genes encode enzymatically active 2-enoyl thioester reductases involved in mitochondrial synthesis of fatty acids (fatty acid synthesis type II) and respiratory competence. The 5′- and 3′-flanking (coding) regions of ETR2 and ETR1 are about 90% (97%) identical, indicating that the genes have evolved via gene duplication. The gene products differ in three amino acid residues: Ile67 (Val), Ala92 (Thr), and Lys251 (Arg) in Etr2p (Etr1p). Quantitative PCR analysis and reverse transcriptase-PCR indicated that both genes were expressed about equally in fermenting and ETR1 predominantly respiring yeast cells. Like the situation with ETR1, expression of ETR2 in respiration-deficient Saccharomyces cerevisiae mutant cells devoid of Ybr026p/Etr1p was able to restore growth on glycerol. Triclosan that is used as an antibacterial agent against fatty acid synthesis type II 2-enoyl thioester reductases inhibited growth of FabI overexpressing mutant yeast cells but was not able to inhibit respiratory growth of the ETR2- or ETR1-complemented mutant yeast cells. Resolving of crystal structures obtained via Etr2p and Etr1p co-crystallization indicated that all possible dimer variants occur in the same asymmetric unit, suggesting that similar dimer formation also takes place in vivo.

KW - Candida tropicalis

KW - gene cloning

KW - genes

KW - reductase

KW - fatty acids

U2 - 10.1074/jbc.M307664200

DO - 10.1074/jbc.M307664200

M3 - Article

VL - 278

SP - 41213

EP - 41220

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 42

ER -