A post-transcriptional mechanism regulates calpastatin expression in bovine skeletal muscle1

Abstract

ABSTRACT:The objective of this study was to investigate whether single nucleotide polymorphisms (SNP) in the calpain 1 (CAPN1), calpain 3 (CAPN3) and calpastatin (CAST) genes, which have been shown to be associated with shear force and tenderness differences in the skeletal muscle of cattle, contribute to phenotypic variation in muscle tenderness by modulating the transcriptional activity of their respective gene. The mRNA expression of the calpain and CAST genes was assessed in the longissimus lumborum muscle (LLM) of cattle from two herds located in distinct production zones on the east (New South Wales, NSW) and west (Western Australia, WA) of Australia. The cattle in the herds were mainly Brahman cattle (Bos indicus) with smaller populations of Angus cattle (Bos taurus). There were 191 steers in the WA herd and 107 steers and 106 heifers in the NSW herd. These herds were established by choosing cattle from the diverse population which had different single nucleotide polymorphism (SNP) genotypes at the CAPN1, CAPN3 and CAST loci. Using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), the transcriptional activities of the CAPN1 and the CAST genes, but not the CAPN3 gene, were found to differ between favorable, positively associated with tenderness, and unfavorable, negatively associated with tenderness, allelic variants of these genes. These findings suggest that the muscle shear force and consumer taste panel differences in tenderness explained by the CAPN1 and CAST gene markers are a consequence of alterations in their mRNA levels, which may ultimately influence the protein activity of these genes, thereby altering the rate and(or) the extent of postmortem proteolysis in skeletal muscle. Of particular importance were the significantly lower type II and type III CAST 5′ splice variant mRNA levels that were detected in the LLM muscle of Brahman and Angus cattle with 2 favourable alleles of the CAST:c.2832A > G polymorphism. Moreover, a reduction in the abundance of an alternative polyadenylated variant of the CAST transcript, terminated at the proximal polyadenylation site, provides a unique insight into the potential involvement of a post-transcriptional regulatory mechanism which may influence protein expression levels in bovine skeletal muscle.

Characterization of the Goose <i>CAPN3</i> Gene and its Expression Pattern in Muscle Tissues of Sichuan White Geese at Different Growth Stages

“…Meanwhile, the rapidity of CAPN3 autodegradation was also thought to through regulate its activity in N-terminal (Ono et al, 2014). Although it has also been demonstrated to be a candidate protein for meat tenderness in mammals, including cattle (Robinson et al, 2012;Nattrass et al, 2014) and pigs (De Smet et al, 2003), its roles, particularly its physiological activities, in geese have not been sufficiently examined. Therefore, this work will have the potential to increase our understanding of the functional roles of CAPN3 gene during improving goose meat quality provides a molecular basis.…”

Differences in skeletal muscle proteolysis in Nellore and Angus cattle might be driven by Calpastatin activity and not the abundance of Calpain/Calpastatin

“…The next question to be answered is what causes the differences in myofibrillar proteolysis in skeletal muscle of Bos taurus and Bos indicus. To answer that question a great body of evidence has been amassed to evaluate the mRNA abundance and/or the allelic as well as genotype frequencies of CAPN1 and CAST and their polymorphism to assess the possible mechanisms related to myofibrillar proteolysis in the skeletal muscle (Curi et al 2009(Curi et al , 2010Giusti et al 2013;Nattrass et al 2014). Although most of these studies were developed to investigate beef tenderness, their results may reflect the effects of these genes on proteolysis in a live skeletal muscle tissue.…”

“…Although most of these studies were developed to investigate beef tenderness, their results may reflect the effects of these genes on proteolysis in a live skeletal muscle tissue. Nattrass et al (2014) speculated that the level of gene expression of CAPN1 and CAST would ultimately influence the activity of the proteins encoded by these genes (Calpain and Calpastatin), thus leading to the difference in the extent of skeletal muscle proteolysis. However, a genome scan performed by Tizioto et al (2013) found a small effect of single nucleotide polymorphisms (SNPs) in CAPN1 and CAST in proteolysis and consequently Warner-Bratzler shear force on beef from Nellore cattle.…”

El sistema proteolítico calpaina en la tenderización de la carne: Un enfoque molecular.

“…Niciura et al (48) found that CAST was expressed twice as much in muscle of homozygous GG as in heterozygous AG in the CAST T1 marker. Natrass et al (49) investigated this relationship in Bos indicus and Bos taurus cattle and found differences in CAPN1 and CAST gene expression between favourable and unfavourable allelic variants of these genes (CAPN1 4751 and CAST T1 markers), indicating a polymorphic effect on gene expression. These findings suggest that differences in tenderness explained by gene markers could be a consequence of the alteration in their mRNA levels, protein activity and rate and (or) the extent of postmortem proteolysis in skeletal muscle.…”