학술논문
Genes encoding the α-carboxyltransferase subunit of acetyl-CoA carboxylase from brassica napus and parental species: cloning, expression patterns, and evolution
Document Type
Report
Author
Source
Genome. May 1, 2010, Vol. 53 Issue 5, p360, 11 p.
Subject
Language
English
ISSN
0831-2796
Abstract
Introduction Brassica napus L. (AACC, 2n = 4x = 38), an amphidiploid species originating from a spontaneous hybridization of B. rapa L. (AA, 2n = 2x = 20) and B. [...]
Heteromeric acetyl coenzyme A carboxylase (ACCase), a rate-limiting enzyme in fatty acid biosynthesis in dicots, is a multi-enzyme complex consisting of biotin carboxylase, biotin carboxyl carrier protein, and carboxyltransferase (α-CT and β-CT). In the present study, four genes encoding α-CT were cloned from Brassica napus, and two were cloned from each of the two parental species, B. rapa and B. oleracea. Comparative and cluster analyses indicated that these genes were divided into two major groups. The major divergence between group-1 and group-2 occurred in the second intron. Group-2 α-CT genes represented the ancestral form in the genus Brassica. The divergence of group-1 and group-2 genes occurred in their common ancestor 12.96-17.78 million years ago (MYA), soon after the divergence of Arabidopsis thaliana and Brassica (15-20 MYA). This time of divergence is identical to that reported for the paralogous subgenomes of diploid Brassica species (13-17 MYA). Real-time reverse transcription PCR revealed that the expression patterns of the two groups of genes were similar in different organs, except in leaves. To better understand the regulation and evolution of α-CT genes, promoter regions from two sets of orthologous gene copies from B. napus, B. rapa, and B. oleracea were cloned and compared. The function of the promoter of gene Bnα-CT-1-1 in group-1 and gene Bnα-CT-2-1 in group-2 was examined by assaying β-glucuronidase activity in transgenic A. thaliana. Our results will be helpful in elucidating the evolution and regulation of ACCase in oilseed rape. Key words: Brassica, acetyl-CoA carboxylase (ACCase), α-CT, evolution. L'aceetyle co-enzyme A carboxylase (ACCase) heteromeerique, une enzyme limitante dans la synthase des acides gras chez les dicotyledones, est un complexe multienzymatique comprenant la biotine carboxylase, la proteine carboxylee transporteuse de biotine et la carboxyltransferase (α-CT et β-CT). Dans ce travail, quatre genes codant pour l'α-CT ont ete clones chez le Brassica napus de meme que deux genes de chacune des deux especes parentales, B. rapa et B. oleracea. Des analyses comparees et de groupement ont indiquee que ces genes forment deux groupes principaux. La divergence majeure entre les groupes 1 et 2 se trouvent au sein du second intron. Les genes du groupe 2 de l'α-CT constituent la forme ancestrale chez le genre Brassica. La divergence des genes des groupes 1 et 2 se serait produite au sein de l'ancetre commun il y a 12,96-17,78 millions d'annees (MA), tot apres la divergence entre l'Arabidopsis thaliana et Brassica (15-20 MA). Ce moment de divergence est identique a celui estimee pour la divergence des sous-genomes paralogues au sein des Brassica diploldes (13-17 MA). Une analyse RT-PCR en temps reel a revele que l'expression des deux groupes de genes est semblable au sein de differents tissus a l'exception des feuilles. Afin de mieux connaitre la regulation et revolution des genes codant pour l'α-CT, les regions promotrices de deux jeux d'orthologues ont ete; cloneees et comparees chez le B. napus,le B. rapa et le B. oleracea. La fonction du promoteur du gene Bnα-CT-1-1 (appartenant au groupe 1) et celui du gene Bnα-CT-2-1 (appartenant au groupe 2) a ete examinee en mesurant l'activite β-glucuronidase chez des lignees transgeniques de l'A. thaliana. Ces resultats seront utiles pour elucider revolution et la regulation de l'ACCase chez le colza okeagineux. Mots-cles : Brassica, acetyl-CoA carboxylase (ACCase), α-CT, evolution. [Traduit par la Redaction]
Heteromeric acetyl coenzyme A carboxylase (ACCase), a rate-limiting enzyme in fatty acid biosynthesis in dicots, is a multi-enzyme complex consisting of biotin carboxylase, biotin carboxyl carrier protein, and carboxyltransferase (α-CT and β-CT). In the present study, four genes encoding α-CT were cloned from Brassica napus, and two were cloned from each of the two parental species, B. rapa and B. oleracea. Comparative and cluster analyses indicated that these genes were divided into two major groups. The major divergence between group-1 and group-2 occurred in the second intron. Group-2 α-CT genes represented the ancestral form in the genus Brassica. The divergence of group-1 and group-2 genes occurred in their common ancestor 12.96-17.78 million years ago (MYA), soon after the divergence of Arabidopsis thaliana and Brassica (15-20 MYA). This time of divergence is identical to that reported for the paralogous subgenomes of diploid Brassica species (13-17 MYA). Real-time reverse transcription PCR revealed that the expression patterns of the two groups of genes were similar in different organs, except in leaves. To better understand the regulation and evolution of α-CT genes, promoter regions from two sets of orthologous gene copies from B. napus, B. rapa, and B. oleracea were cloned and compared. The function of the promoter of gene Bnα-CT-1-1 in group-1 and gene Bnα-CT-2-1 in group-2 was examined by assaying β-glucuronidase activity in transgenic A. thaliana. Our results will be helpful in elucidating the evolution and regulation of ACCase in oilseed rape. Key words: Brassica, acetyl-CoA carboxylase (ACCase), α-CT, evolution. L'aceetyle co-enzyme A carboxylase (ACCase) heteromeerique, une enzyme limitante dans la synthase des acides gras chez les dicotyledones, est un complexe multienzymatique comprenant la biotine carboxylase, la proteine carboxylee transporteuse de biotine et la carboxyltransferase (α-CT et β-CT). Dans ce travail, quatre genes codant pour l'α-CT ont ete clones chez le Brassica napus de meme que deux genes de chacune des deux especes parentales, B. rapa et B. oleracea. Des analyses comparees et de groupement ont indiquee que ces genes forment deux groupes principaux. La divergence majeure entre les groupes 1 et 2 se trouvent au sein du second intron. Les genes du groupe 2 de l'α-CT constituent la forme ancestrale chez le genre Brassica. La divergence des genes des groupes 1 et 2 se serait produite au sein de l'ancetre commun il y a 12,96-17,78 millions d'annees (MA), tot apres la divergence entre l'Arabidopsis thaliana et Brassica (15-20 MA). Ce moment de divergence est identique a celui estimee pour la divergence des sous-genomes paralogues au sein des Brassica diploldes (13-17 MA). Une analyse RT-PCR en temps reel a revele que l'expression des deux groupes de genes est semblable au sein de differents tissus a l'exception des feuilles. Afin de mieux connaitre la regulation et revolution des genes codant pour l'α-CT, les regions promotrices de deux jeux d'orthologues ont ete; cloneees et comparees chez le B. napus,le B. rapa et le B. oleracea. La fonction du promoteur du gene Bnα-CT-1-1 (appartenant au groupe 1) et celui du gene Bnα-CT-2-1 (appartenant au groupe 2) a ete examinee en mesurant l'activite β-glucuronidase chez des lignees transgeniques de l'A. thaliana. Ces resultats seront utiles pour elucider revolution et la regulation de l'ACCase chez le colza okeagineux. Mots-cles : Brassica, acetyl-CoA carboxylase (ACCase), α-CT, evolution. [Traduit par la Redaction]