Cloning, comparative characterization of porcine SCAP gene, and identification of its two splice variants

Abstract

Sterol responsive element binding protein (SREBP) cleavage-activating protein (SCAP) is the key regulator of activation of SREBPs, which stimulate most enzymes in cholesterol and lipid synthesis. In order to investigate the molecular basis of lipid metabolism in the pig, a unique model for fat deposition, we isolated and characterized the porcine SCAP. The 4,096-bp full-length porcine SCAP cDNA contains an open reading frame of 3,840 bp. The predicted SCAP protein consists of 1,280 amino acids of 55-92% identity with its vertebrate counterparts. The porcine SCAP gene consists of at least 19 exons and 18 introns, which span over 13 kb of the genome. The porcine SCAP gene was mapped to chromosome 13q21-22 using a porcine-rodent somatic cell hybrid panel. Comparison of SCAP genomic structures from various species revealed intron losses in porcine, Tetraodon and fugu SCAP, and intron gains in cow and chicken SCAP. Moreover, we isolated two novel splicing SCAP variants with 193-bp (variant 2) in-frame deletion from testis and a variant with 291-bp (variant 3) in-frame deletion from liver and muscle, which may affect the function of the porcine SCAP. In conclusion, the intron gains and losses appear to have contributed to the shape of the modern SCAP family. The splice variants detected, first to be reported in any species, may be involved in the particulars of the fat metabolism in the pig. Our data lay foundation for further study of SCAP function in this species.

SREBP pathway genes as candidate markers in country ham production

“…Initially, primer sets were used for each of these genes in order to amplify PCR fragments from each gene. SREBF1 primer set (Table 1) was designed from the corresponding human SREBF1 exon 6 genomic sequence, SCAP primer set was described in Qiu et al, (2006) (Forward primer: GCAGCGC-CGGGACAGTGG; reverse primer: TGATGCCC-GAGGCCACCTCCTC) and amplified intron 13 of the SCAP gene. MBTPS1 primers were designed from the consensus regions of exon 5 (Forward primer: TGAAGGAGAGAACCAACTG-GA) and exon 6 (reverse primer: CTGCATCTG-GAGCAAATCCT) of the human (accession numbers in ensemble ENSE00000945694 and ENSE00000945695) and mouse (accession numbers: ENSMUSE00000319523 and ENS-MUSE00000319518) MBTPS1 gene.…”

Molecular characterisation of the buffaloSCAPgene and its association with milk production traits in water buffaloes

“…SREBP cleavage-activating protein ( SCAP ) is a family of endoplasmic reticulum (ER) membrane-bound transcription factors required for cleavage and activation of sterol regulatory element-binding proteins (SREBPs), which play a critical role in the regulation of cholesterol homeostasis (Weber et al 2004). To date, SCAP gene has been cloned and characterised from several vertebrate (Qiu et al 2006) and non-vertebrate species (Seegmiller et al 2002). Several studies have confirmed that the single nucleotide polymorphisms (SNPs) in SCAP might affect intracellular lipid metabolism (Iwaki et al 1999), metabolic phenotypes (Cao et al 2002), and obesity (Liu et al 2014).…”