Full Length cDNA Molecular Cloning of Acetyl-CoA Carboxylase &alpha; Gene and Effects of Dietary Lipid Level on Its Expression in the Liver of Half-Smooth Tongue Sole (Cynoglossus semilaevis)

ZHANG Xiaqing; XU Jianhe; PAN Qian; YI Lefei; PENG Yongxing; SHEN Xin; YAN Binlun; GAO Huan; WANG Wenxiang; CHENG Hanliang

doi:10.3969/j.issn.1006-267x.2016.02.022

2016 , Vol. 28 >Issue 2: 485 - 497

DOI: https://doi.org/10.3969/j.issn.1006-267x.2016.02.022

Molecular Nutrition

Full Length cDNA Molecular Cloning of Acetyl-CoA Carboxylase α Gene and Effects of Dietary Lipid Level on Its Expression in the Liver of Half-Smooth Tongue Sole (Cynoglossus semilaevis)

ZHANG Xiaqing ,
XU Jianhe ,
PAN Qian ,
YI Lefei ,
PENG Yongxing ,
SHEN Xin ,
YAN Binlun ,
GAO Huan ,
WANG Wenxiang ,
CHENG Hanliang

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1. Huaihai Institute of Technology, Lianyungang 222005, China;
2. Co-Innovation Center of Jiangsu Marine Bio-Industry Technology, Lianyungang 222005, China;
3. Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China

Received date: 2015-08-01

Online published: 2016-02-19

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Abstract

The full-length cDNA of acetyl-coa carboxylase α (ACC1) gene was cloned from liver of half-smooth tongue sole (Cynoglossus semilaevis) by reverse transcription PCR (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. The expression of ACC1 mRNA in gut, liver, muscle, ovary, spleen, brain, kidney, heart, mesenteric adipose tissue of half-smooth tongue sole was analyzed by real-time fluorescence quantitative PCR (RT-qPCR) method. In addition, the effects of dietary lipid level on ACC1 gene expression in liver of half-smooth tongue sole were investigated. The results showed as follows:1) the full-length cDNA of ACC1 gene was 7 811bp with a 7 074 bp open reading frame encoding 2 357 amino acids. The ACC1 protein has a calculated molecular weight of 266 ku and isolectric point of 6.42. Some conserved sites of ACC1 amino acid sequence were found, including a ATP-binding site (Gly³¹⁶ to Gly³²¹), a biotin-binding site (Val⁷⁸⁵-Met⁷⁸⁶-Lys⁷⁸⁷-Met⁷⁸⁸), and a CoA-binding site (Ser¹⁹⁶⁹ to Val¹⁹⁹⁵). In addition, the alternative splice varieties were found in ACC1 gene, and we present evidence for the presence of two isoforms of ACC1 in half-smooth tongue sole liver that differ from the 266 ku ACC1 by the absence of 8 and 15 amino acids. 2) The expression of ACC1 mRNA was detected in all examined tissues. The relative expression level of ACC1 mRNA in liver and brain were 2.67 and 2.53, respectively, which were significantly higher than that in other tissues (P<0.05); the relative expression level of ACC1 mRNA in gut, mesenteric adipose and ovary were second, which were 1.14, 1.10 and 0.97, respectively; the relative expression level of ACC1 mRNA in kidney was the lowest, only was 0.48. 3) Compared with the control group (without fish oil group), diet added with 3.5% fish oil significantly decreased the relative expression level of ACC1 mRNA in liver (P<0.05); diet added with 7.0% and 10.0% fish oil led to further decrease the relative expression level of ACC1 mRNA in liver, and it was significantly lower than that in 3.5% fish oil group and control group (P<0.05), meanwhile, the 10.0% fish oil group was lower than 7.0% fish oil group (P>0.05). In summary, we have cloned the full-length cDNA of ACC1 gene from half-smooth tongue sole. Compared with other vertebrates, the main functional sites (ATP-binding site, biotin-binding site and CoA-binding site) are basically conserved. The lipogenic tissues, such as liver and brain, are the main ACC1 gene expressing tissues in half-smooth tongue sole. Moreover, liver ACC1 mRNA expression is inhibited after the fish fed diets added with fish oil, and the inhibitory effect is positively correlated with the addition of fish oil.

Key words： half-smooth tongue sole (Cynoglossus semilaevis); ACC1; molecular cloning; nutritional regulation; tissue express

Cite this article

ZHANG Xiaqing , XU Jianhe , PAN Qian , YI Lefei , PENG Yongxing , SHEN Xin , YAN Binlun , GAO Huan , WANG Wenxiang , CHENG Hanliang . Full Length cDNA Molecular Cloning of Acetyl-CoA Carboxylase α Gene and Effects of Dietary Lipid Level on Its Expression in the Liver of Half-Smooth Tongue Sole (Cynoglossus semilaevis)[J]. Chinese Journal of Animal Nutrition, 2016 , 28(2) : 485 -497 . DOI: 10.3969/j.issn.1006-267x.2016.02.022

References

[1] BARBER M C,PRICE N T,TRAVERS M T.Structure and regulation of acetyl-CoA carboxylase genes of metazoa[J]. Biochimica et Biophysica Acta:Molecular and Cell Biology of Lipids,2005,1733(1):1-28.

[2] TONG L.Acetyl-coenzyme a carboxylase:crucial metabolic enzyme and attractive target for drug discovery[J].Cellular and Molecular Life Sciences,2005,62(16):1784-1803.

[3] JITRAPAKDEE S,WALLACE J C.The biotin enzyme family:conserved structural motifs and domain rearrangements[J].Current Protein & Peptide Science,2003,4(3):217-219.

[4] LÓPEZ-CASILLAS F,BAI D H,LUO X C,et al.Structure of the coding sequence and primary amino acid sequence of acetyl-coenzyme A carboxylase[J].Proceedings of the National Academy of Sciences of the United States of America,1988,85(16):5784-5788.

[5] TAKAI T,YOKOYAMA C,WADA K,et al.Primary structure of chicken liver acetyl-CoA carboxylase deduced from cDNA sequence[J].Journal of Biological Chemistry,1988,263(6):2651-2657.
[6] ABU-ELHEIGA L,ALMARZA-ORTEGA D B,BALDINI A,et al.Human acetyl-CoA carboxylase 2 Molecular cloning,characterization,chromosomal mapping,and evidence for two isoforms[J].Journal of Biological Chemistry,1997,272(16):10669-10677.

[7] ABU-ELHEIGA L,JAYAKUMAR A,BALDINI A,et al.Human acetyl-CoA carboxylase:characterization,molecular cloning,and evidence for two isoforms[J].Proceedings of the National Academy of Sciences of the United States of America,1995,92(9):4011-4015.

[8] ZHANG H L,YANG Z R,SHEN Y,et al.Crystal structure of the carboxyltransferase domain of acetyl-coenzyme A carboxylase[J].Science,2003,299(5615):2064-2067.

[9] MAO J,MARCOS S,DAVIS S K,et al.Genomic distribution of three promoters of the bovine gene encoding acetyl-CoA carboxylase alpha and evidence that the nutritionally regulated promoter Ⅰ contains a repressive element different from that in rat[J].Biochemical Journal,2001,358(Pt.1):127-135.
[10] BARBER M C,TARVERS M T.Cloning and characterisation of multiple acetyl-CoA carboxylase transcripts in ovine adipose tissue[J].Gene,1995,154(2):271-275.

[11] BADAOUI B,SERRADILLA J M,TOMÀS A,et al.Goat acetyl-coenzyme a carboxylase α:molecular characterization,polymorphism,and association with milk traits[J].Journal of Dairy Science,2007,90(2):1039-1043.

[12] CHENG H L,JI N J,PENG Y X,et al.Molecular characterization and tissue-specific expression of the acetyl-CoA carboxylase α gene from grass carp,Ctenopharyngodon idella[J].Gene,2011,487(1):46-51.

[13] DIEFFENBACH C W,DVEKSLER G S.PCR primer:a laboratory manual[M].Plainview NY:Cold Spring Harbor Laboratory Press,1995.
[14] TAMURA K,STECHER G,PETERSON D,et al.MEGA6:molecular evolutionary genetics analysis version 6.0[J].Molecular Biology and Evolution,2013,30(12):2725-2729.

[15] JITRAPAKDEE S,ST MAURICE M,RAYMENT I,et al.Structure,mechanism and regulation of pyruvate carboxylase[J].Biochemical Journal,2008,413(3):369-387.

[16] LEE C K,CHEONG H K,RYU K S,et al.Biotinoyl domain of human acetyl-CoA carboxylase:structural insights into the carboxyl transfer mechanism[J].Proteins:Structure,Function,and Bioinformatics,2008,72(2):613-624.

[17] BAI D H,MOON T W,LOPEZ-CASILLAS F,et al.Analysis of the biotin-binding site on acetyl-CoA carboxylase from rat[J].European Journal of Biochemistry,1989,182(2):239-245.

[18] BARBER M C,POOLEY L,TRAVERS M T.Developmental regulation of alternatively spliced acetyl-CoA carboxylase-alpha mRNAs encoding isozymes with or without an eight amino acid domain upstream of the Ser-1200 phosphorylation motif in the mammary gland[J].Journal of Molecular Endocrinology,2001,27(3):349-356.

[19] SALLES J,SARGUEIL F,KNOLL-GELLIDA A,et al.Acetyl-CoA carboxylase and SREBP expression during peripheral nervous system myelination[J].Biochimica et Biophysica Acta:Molecular and Cell Biology of Lipids,2003,1631(3):229-238.

[20] KOO H Y,WALLING M A,CHUNG B H,et al.Dietary fructose induces a wide range of genes with distinct shift in carbohydrate and lipid metabolism in fed and fasted rat liver[J].Biochimica et Biophysica Acta:Molecular Basis of Disease,2008,1782(5):341-348.

[21] TAI C C,DING S T.N-3 polyunsaturated fatty acids regulate lipid metabolism through several inflammation mediators:mechanisms and implications for obesity prevention[J].Journal of Nutritional Biochemistry,2010,21(5):357-363.

[22] KAJIKAWA S,HARADA T,KAWASHIMA A,et al.Highly purified eicosapentaenoic acid prevents the progression of hepatic steatosis by repressing monounsaturated fatty acid synthesis in high-fat/high-sucrose diet-fed mice[J].Prostaglandins,Leukotrienes and Essential Fatty Acids,2009,80(4):229-238.

[23] 马晶晶,邵庆均,许梓荣,等.n-3高不饱和脂肪酸对黑鲷幼鱼生长及脂肪代谢的影响[J].水产学报,2009,33(4):639-649.
[24] 李超.n-3高不饱和脂肪酸对草鱼生长、脂代谢及健康状况的影响[D].硕士学位论文.杨凌:西北农林科技大学,2013.
[25] ZHOU J C,HAN D,JIN J Y,et al.Compared to fish oil alone,a corn and fish oil mixture decreases the lipid requirement of a freshwater fish species,Carassius auratus gibelio[J].Aquaculture,2014,428/429:272-279.

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