Abstract

Homocysteine can be converted to cysteine via the transsulfuration pathway where cystathionine beta-synthase (CBS) is a rate-limiting enzyme. Homocysteine is thought to play a role in bone remodelling and strength. Previous results indicated that some of the difference in bone strength of end-of-lay hens may be associated with CBS expression level. To investigate if differences in the promoter region of the CBS gene might be responsible for observed differences in gene expression between CBS alleles post mortem- and in-vitro expression studies have been undertaken. Transfection of the DF-1 avian cell line with a series of deletion fragments of the 5' promoter, or constructs containing three CBS allele sequences, with a luciferase reporter revealed that a core region of -155 to + 131 bp in the CBS promoter was essential for mRNA expression. We found that a variable number of short tandem repeats (7 nucleotide motif and 6 nucleotide repeats) in the core region of the promoter affecting the transcriptional activity and a strong effect for gene expression. However, methylation of the 6 nucleotide repeats varied between allelic variants and these maybe responsible for differences in promoter activity. Our findings indicated variable short tandem repeats and the differentially methylated sites in the promoter region may be responsible for CBS expression differences in the bone of laying hens.

Keywords

Cystathionine beta-synthase; Laying hen; Methylation; Promoter; Short tandem repeat

Published in

Gene
2019, Volume: 710, pages: 367-374
Publisher: ELSEVIER

SLU Authors

• de Koning, Dirk-Jan

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Genetics and Breeding


Publication identifier

DOI: https://doi.org/10.1016/j.gene.2019.05.049

Permanent link to this page (URI)

https://res.slu.se/id/publ/101653