1 Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountains Region of Ministry of Education (Guizhou University)/Key Laboratory of Animal Genetics, Breeding and Reproduction in Guizhou, Guiyang 550025, China;
2 College of Animal Sciences, Guizhou University, Guiyang 550025, China;
3 College of Life Sciences, Guizhou University, Guiyang 550025, China
Abstract：PRKAA1 (protein kinase, AMP activated α1) plays a very important regulatory role for the processes of fat metabolism, cell proliferation, fat formation, etc. In this study, Guizhou fatty pig breed Congjiang Xiang pig (Sus scrofa) and commercial pig breed Yorkshire pigs (S. scrofa) were taken as research objects. Based on the 3'UTR sequence of PRKAA1 (GenBank No.XM_021076521), NCBI and NONCODE (http://www.noncode.org/) were used to screen the LncRNAs related to PRKAA1. The screened LncRNAs were cloned, and the secondary structure, ORF, target site of PRKAA1 and regulated miRNAs were predicted. qRT-PCR was used to detect the screened LncRNAs expression in different tissues of Congjiang Xiang pigs and Yorkshire pigs. Two candidate LncRNAs, NONSUST030689.1 (named ALNC1) and NONSUST004358.1 (named ALNC2), were selected for next experiments. The sequenced results revealed that ALNC2 had 46 bp inserted compared with original sequence. ALNC1 had 4 ORFs, and ALNC2 had 5 ORFs. In addition, ALNC1 and ALNC2 each had 1 target site for PRKAA1. Predicted miRNAs which could bind with LncRNAs including miR-1226-5p, miR-487b-5p, miR-7156-5p, etc. The results of qRT-PCR showed that the 2 LncRNAs had universal expression in all tissues of Congjiang Xiang pig and Yorkshire pig and were significantly different among different tissues. In Congjiang Xiang pig, ALNC1 and ALNC2 were the highest expressed in the liver and spleen and the lowest in the longissimus dorsi muscle. But ALNC1 and ALNC2 were the highest expressed in the large intestine and lung and the lowest in the longissimus dorsi muscle in Yorkshire pig. ALNC1 had significant higher expression in liver, spleen, small intestine, fat, and longissimus dorsi muscle of Congjiang Xiang pigs than that of Yorkshire pigs (P<0.01), but lower expression in heart and large intestine than that of Yorkshire pigs (P<0.01), and no significant difference in kidney and lung. ALNC2 had significant higher expression in liver, kidney and longissimus dorsi muscle of Congjiang Xiang pigs than that of Yorkshire pigs (P<0.01), but lower expression in heart, lung, small intestine, and large intestine (P<0.01). In fat tissues, ALNC2 expression levels of Congjiang Xiang pig were significantly lower than that of Yorkshire pigs (P<0.05). The ALNC2 expression levels in spleen had no significant difference between Congjiang Xiang pigs and Yorkshire pigs. The present results could provide basic data for further studies on the regulatory role of PRKAA1 and LncRNAs in fat deposition process of pigs.
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