Browse > Article
http://dx.doi.org/10.5713/ajas.17.0911

Novel splice isoforms of pig myoneurin and their diverse mRNA expression patterns  

Guo, Xiaohong (College of Animal Science and Veterinary Medicine, Shanxi Agricultural University)
Li, Meng (College of Animal Science and Veterinary Medicine, Shanxi Agricultural University)
Gao, Pengfei (College of Animal Science and Veterinary Medicine, Shanxi Agricultural University)
Cao, Guoqing (College of Animal Science and Veterinary Medicine, Shanxi Agricultural University)
Cheng, Zhimin (College of Animal Science and Veterinary Medicine, Shanxi Agricultural University)
Zhang, Wanfeng (College of Animal Science and Veterinary Medicine, Shanxi Agricultural University)
Liu, Jianfeng (Key Laboratory of Animal Genetics Breeding and Reproduction, Ministry of Agriculture, College of Animal Science and Technology, China Agricultural University)
Liu, Xiaojun (College of Animal Science and Veterinary Medicine, Henan Agricultural University)
Li, Bugao (College of Animal Science and Veterinary Medicine, Shanxi Agricultural University)
Publication Information
Asian-Australasian Journal of Animal Sciences / v.31, no.10, 2018 , pp. 1581-1590 More about this Journal
Abstract
Objective: The aim of this study was to clone alternative splicing isoforms of pig myoneurin (MYNN), predict the structure and function of coding protein, and study temporal and spatial expression characteristics of each transcript. Methods: Alternative splice isoforms of MYNN were identified using RNA sequencing (RNA-seq) and cloning techniques. Quantitative real-time polymerase chain reaction (qPCR) was employed to detect expression patterns in 11 tissues of Large White (LW) and Mashen (MS) pigs, and to study developmental expression patterns in cerebellum (CE), stomach (ST), and longissimus dorsi (LD). Results: The results showed that MYNN had two alternatively spliced isoforms, MYNN-1 (GenBank accession number: KY470829) and MYNN-2 (GenBank accession number: KY670835). MYNN-1 coding sequence (CDS) is composed of 1,830 bp encoding 609 AA, whereas MYNN-2 CDS is composed of 1,746 bp encoding 581 AA. MYNN-2 was 84 bp less than MYNN-1 and lacked the sixth exon. MYNN-2 was found to have one $C_2H_2$ type zinc finger protein domain less than MYNN-1. Two variants were ubiquitously expressed in all pig tissues, and there were significant differences in expression of different tissues (p<0.05; p<0.01). The expression of MYNN-1 was significantly higher than that of MYNN-2 in almost tissues (p<0.05; p<0.01), which testified that MYNN-1 is the main variant. The expression of two isoforms decreased gradually with increase of age in ST and CE of MS pig, whereas increased gradually in LW pig. In LD, the expression of two isoforms increased first and then decreased with increase of age in MS pig, and decreased gradually in LW pig. Conclusion: Two transcripts of pig MYNN were successfully cloned and MYNN-1 was main variant. MYNN was highly expressed in ST, CE, and LD, and their expression was regular. We speculated that MYNN plays important roles in digestion/absorption and skeletal muscle growth, whereas the specific mechanisms require further elucidation.
Keywords
Pig; MYNN; Alternative Splicing; mRNA Expression;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Aravind L, Koonin EV. Fold prediction and evolutionary analysis of the POZ domain: structural and evolutionary relationship with the potassium channel tetramerization domain. J Mol Biol 1999;285:1353-61.   DOI
2 Li X, Peng H, Schultz DC, et al. Structure-function studies of the BTB/POZ transcriptional repression domain from the promyelocytic leukemia zinc finger oncoprotein. Cancer Res 1999;59:5275-82.
3 Takenaga M, Hatano M, Takamori M, et al. Bcl6-dependent transcriptional repression by BAZF. Biochem Biophys Res Commun 2003;303:600-8.   DOI
4 Phan RT, Dalla-Favera R. The BCL6 proto-oncogene suppresses p53 expression in germinal-centre B cells. Nature 2004;432:635-9.   DOI
5 Cifuentes-Diaz C, Bitoun M, Goudou D, et al. Neuromuscular expression of the BTB/POZ and zinc finger protein myoneurin. Muscle Nerve 2004;29:59-65.   DOI
6 Alliel PM, Seddiqi N, Goudou D, et al. Myoneurin, a novel member of the BTB/POZ-zinc finger family highly expressed in human muscle. Biochem Biophys Res Commun 2000;273:385-91.   DOI
7 Alliel PM, Cifuentes-Diaz C, Bitoun M, et al. Expression of the transcriptional repressor gene myoneurin at the neuromuscular junction, during mouse development and in the adult. J Physiol Paris 2006;99:1.
8 Prag S, Collett GD, Adams JC. Molecular analysis of muskelin identifies a conserved discoidin-like domain that contributes to protein self-association. Biochem J 2004;381:547-59.   DOI
9 Kelly KF, Otchere AA, Graham M, Daniel JM. Nuclear import of the BTB/POZ transcriptional regulator Kaiso. J Cell Sci 2004;117:6143-52.   DOI
10 Huynh KD, Bardwell VJ. The BCL-6 POZ domain and other POZ domains interact with the co-repressors N-CoR and SMRT. Oncogene 1998;17:2473-84.   DOI
11 Li X, Zhang D, Hannink M, Beamer LJ. Crystal structure of the Kelch domain of human Keap1. J Biol Chem 2004;279:54750-8.   DOI
12 Ledee DR, Gao CY, Seth R, et al. A specific interaction between muskelin and the cyclin-dependent kinase 5 activator p39 promotes peripheral localization of muskelin. J Biol Chem 2005;280:21376-83.   DOI
13 Melnick A, Carlile G, Ahmad KF, et al. Critical residues within the BTB domain of PLZF and Bcl-6 modulate interaction with corepressors. Mol Cell Biol 2002;22:1804-18.   DOI
14 David G, Alland L, Hong SH, et al. Histone deacetylase associated with mSin3A mediates repression by the acute promyelocytic leukemia-associated PLZF protein. Oncogene 1998;16:2549-56.   DOI
15 Ahmad KF, Melnick A, Lax S, et al. Mechanism of SMRT corepressor recruitment by the BCL6 BTB domain. Mol Cell 2003;12:1551-64.   DOI
16 Zhou C, Li LY. Advances in C2H2 zinc finger proteins. Life Sci Res 2004;8:215-20.
17 Mitchelmore C, Kjaerulff KM, Pedersen HC, et al. Characterization of two novel nuclear BTB/POZ domain zinc finger isoforms. Association with differentiation of hippocampal neurons, cerebellar granule cells, and macroglia. J Biol Chem 2002;277:7598-609.   DOI
18 Meng YY, Xu XZ, Liao J. ZBTB transcription factors and tumorigenesis. Life Sci 2015;27:477-85.
19 Trappe R, Buddenberg P, Uedelhoven J, et al. The murine BTB/POZ zinc finger gene Znf131: predominant expression in the developing central nervous system, in adult brain, testis, and thymus. Biochem Biophys Res Commun 2002;296:319-27.   DOI
20 Sakashita C, Fukuda T, Okabe S, et al. Cloning and characterization of the human BAZF gene, a homologue of the BCL6 oncogene. Biochem Biophys Res Commun 2002;291:567-73.   DOI
21 Zhao YY, Gao PF, Li W, et al. Study on the developmental expression of Lbx1 gene in longissimus dorsi of Mashen and Large White Pigs. Italian J Anim Sci 2015;14:2081-7.
22 Trapnell C, Pachter L, Salzberg SL. TopHat: discovering splice junctions with RNA-Seq. Bioinformatics 2009;25:1105-11.   DOI
23 Marquez Y, Brown JW, Simpson C, Barta A, Kalyna M. Transcriptome survey reveals increased complexity of the alternative splicing landscape in Arabidopsis. Genome Res 2012;22:1184-95.   DOI
24 Graveley BR, Brooks AN, Carlson JW, et al. The developmental transcriptome of Drosophila melanogaster. Nature 2011;471:473-9.   DOI
25 Mortazavi A, Williams BA, McCue K, Schaeffer L, Wold B. Mapping and quantifying mammalian transcriptomes by RNA-Seq. Nat Methods 2008;5:621-8.   DOI
26 Ghosh S, Chan CK. Analysis of RNA-Seq data using TopHat and Cufflinks. Methods Mol Biol 2016;1374:339-61.
27 Zhang J. Species diversity and the way to protect MA SHEN ZHU. Shanxi, China: J Shanxi Agric Univ; 2001.
28 Yang WP, Cao GQ, Shi JZ, Liu JH, Zhou ZX. Study on the finishing ability of different cross combination in pig. Chinese J Anim Sci 2005;41:48-9.
29 Kim D, Pertea G, Trapnell C, et al. TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol 2013;14:R36.   DOI
30 Zhang XN, Xiao HS. Progress on the method of identification of alternative splicing. Life Science Instruments; 2008.
31 Modrek B, Lee C. A genomic view of alternative splicing. Nat Genet 2002;30:13-9.   DOI
32 Niu JY. Study on the developmental expression of Pax7, MyoD and MyoG genes in longissimus dorsi of pigs. Shanxi, China: Shanxi Agricultural University; 2015.
33 Feng H, Qin Z, Zhang X. Opportunities and methods for studying alternative splicing in cancer with RNA-Seq. Cancer Lett 2013;340:179-91.   DOI
34 Ashmore CR, Addis PB, Doerr L. Development of muscle fibers in the fetal pig. J Anim Sci 1973;36:1088-93.   DOI