Cloning and Expression Analysis of YTHDF2 in Yak (Bos grunniens)
MA Lan-Hua1,2, ZHANG Yong-Feng1, GU Ya-Rong1,2, CHEN Yi-Wei1,2, YAN Ping1,2,*, PAN He-Ping2,*
1 Key Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Animal Husbandry and Veterinary Drug, Chinese Academy of Agricultural Sciences, Lanzhou 730030, China; 2 Life Science and Engineering College, Northwest Minzu University, Lanzhou 730030, China
Abstract：As RNA-binding protein, YT521-b homology domains 2 (YTHDF2) can recognize m6A modifications and plays a pivotal role in mammalian growth and development. In order to reveal the role of yak (Bos grunniens) YTHDF2 gene in yak fat deposition, the CDS of YTHDF2 mRNA was cloned for bioinformatics analysis in perirenal adipose tissue of yak, and the spatiotemporal expression pattern of YTHDF2 mRNA was analyzed by qPCR. The result showed that the total length of YTHDF2 mRNA CDS region was 1 743 bp, encoding 580 amino acids; YTHDF2 was most closely related to wild yak; the amino acid isoelectric point was 8.87, and the instability coefficient (Ⅱ) was 50.00, grand average of hydropathicity was -0.66, YTHDF2 showed strong hydrophilicity in general, YHDF2 had a conserved domain YTH of 133 bp, no signal peptide and transmembrane structural domain; there were 69 potential phosphorylation sites. The advanced structure of YTHDF2 protein was composed of irregularly coiled (67.59%), α-helix (15.15%), extended chain (12.24%) and β-turn (4.66%) connected with each other. YTHDF2 interacted with methylation modification-related proteins and played an important role in RNA specific binding. The results of qPCR showed that YTHDF2 gene was expressed in all 7 tissues of yak, with the highest expression in the longest dorsal muscle (P<0.05); In terms of time, there was no significant difference in the expression level of YTHDF2 at 18 and 30 months in adipose tissue; Furthermore, YTHDF2 expression showed an increasing trend during the differentiation of prerenal adipocytes, and the expression of YTHDF2 at 4, 8 and 12 d was significantly higher than that 0 d (P<0.05). The results of this study showed that YTHDF2 was an unstable alkaline water-soluble protein with a YTH conserved domain, which initially indicated that YTHDF2 gene played an important role in the process of adipose deposition in yaks. This study provides a theoretical basis for further study on the regulation and function of YTHDF2 gene on adipose deposition of yak.
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