Molecular characterization, tissue expression and nucleotide variation of the porcine AZ1 gene

Abstract

Antizyme 1 (AZ1) is a member of the antizyme family that is involved in many biological processes. As a natural inhibitor, AZ1 controls the normal level of polyamines, which is indispensable to cellular function. Our prior research showed that the expression of the AZ1 gene in Longissimus doris of Landrace pigs was higher (P<0.05) than in Lantang pigs. The AZ1 gene might be involved in the development of muscle and potentially serves as an important target for muscle improvement in pigs. In this study, the molecular characterization, tissue expression, and sequence variation of porcine AZ1 gene were analyzed. A 4082 bp sequence including the 5'-flanking region and gene sequence was obtained through RACE and sequencing. The genomic sequence of AZ1 gene consists of six exons and five introns. The mRNA of AZ1 gene contains three elements: 5'-untranslated regions (UTR, 79 bp), CDS (684 or 207 bp), and 3'-UTR (161 or 423 bp for the two transcripts). Three termination signals of AATAAA were found in the longer 3'-UTR. The mRNA sequence of the AZ1 gene contained two ORFs with a frameshifting site at the 69th codon. The amino acid sequence from the porcine AZ1 gene was similar to other vertebrates and exhibited the highest similarity to cattle. The partial 5'-flanking region was 852 bp with typical cis-regulatory elements such as TATA boxes and CAAT boxes. By DNA pooling and sequencing, nine and 12 single nucleotide polymorphisms (SNPs) were detected in the 5'-flanking region and introns, respectively. Except the SNP at -713, the other eight SNPs were found in putative cis-acting elements and might alter the binding of trans-acting factors. Expression patterns showed that in Lantang pig, the AZ1 gene is differentially expressed in various tissues and displayed higher expression in lung and skeletal muscle.