Identification of the Core Promoter and Variants Regulating Chicken <i>CCKAR</i> Expression
Decreased expression of chicken cholecystokinin A receptor (<i>CCKAR</i>) attenuates satiety, which contributes to increased food intake and growth for modern broilers. The study aims to define the core promoter of <i>CCKAR</i>, and to identify variants associated with expres...
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| Format: | Article |
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MDPI AG
2022-06-01
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| Series: | Genes |
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| Online Access: | https://www.mdpi.com/2073-4425/13/6/1083 |
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| author | Zhepeng Wang Angus M. A. Reid Peter W. Wilson Ian C. Dunn |
| author_facet | Zhepeng Wang Angus M. A. Reid Peter W. Wilson Ian C. Dunn |
| author_sort | Zhepeng Wang |
| collection | DOAJ |
| description | Decreased expression of chicken cholecystokinin A receptor (<i>CCKAR</i>) attenuates satiety, which contributes to increased food intake and growth for modern broilers. The study aims to define the core promoter of <i>CCKAR</i>, and to identify variants associated with expression activity. A 21 kb region around the <i>CCKAR</i> was re-sequenced to detect sequence variants. A series of 5′-deleted promoter plasmids were constructed to define the core promoter of <i>CCKAR</i>. The effects of sequence variants located in promoter (PSNP) and conserved (CSNP) regions on promoter activity were analyzed by comparing luciferase activity between haplotypes. A total of 182 variants were found in the 21 kb region. There were no large structural variants around <i>CCKAR</i>. pNL−328/+183, the one with the shortest insertion, showed the highest activity among the six promoter constructs, implying that the key cis elements regulating <i>CCKAR</i> expression are mainly distributed 328 bp upstream. We detected significant activity differences between high- and low-growth associated haplotypes in four of the six promoter constructs. The high-growth haplotypes of constructs pNL−1646/+183, pNL−799/+183 and pNL−528/+183 showed lower activities than the low-growth haplotypes, which is consistent with decreased expression of <i>CCKAR</i> in high-growth chickens. Lower expression of the high-growth allele was also detected for the CSNP5-containing construct. The data suggest that the core promoter of <i>CCKAR</i> is located the 328 bp region upstream from the transcription start site. Lower expression activities shown by the high-growth haplotypes in the reporter assay suggest that CSNP5 and variants located between 328 bp and 1646 bp upstream form a promising molecular basis for decreased expression of <i>CCKAR</i> and increased growth in chickens. |
| first_indexed | 2024-03-09T23:42:49Z |
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| id | doaj.art-56d586f31b274e7981255653ea608082 |
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| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-09T23:42:49Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-56d586f31b274e7981255653ea6080822023-11-23T16:49:00ZengMDPI AGGenes2073-44252022-06-01136108310.3390/genes13061083Identification of the Core Promoter and Variants Regulating Chicken <i>CCKAR</i> ExpressionZhepeng Wang0Angus M. A. Reid1Peter W. Wilson2Ian C. Dunn3College of Animal Science and Technology, Northwest A&F University, Yangling 712100, ChinaRoyal (Dick) School of Veterinary Studies, Roslin Institute, University of Edinburgh, Midlothian EH25 9RG, UKRoyal (Dick) School of Veterinary Studies, Roslin Institute, University of Edinburgh, Midlothian EH25 9RG, UKRoyal (Dick) School of Veterinary Studies, Roslin Institute, University of Edinburgh, Midlothian EH25 9RG, UKDecreased expression of chicken cholecystokinin A receptor (<i>CCKAR</i>) attenuates satiety, which contributes to increased food intake and growth for modern broilers. The study aims to define the core promoter of <i>CCKAR</i>, and to identify variants associated with expression activity. A 21 kb region around the <i>CCKAR</i> was re-sequenced to detect sequence variants. A series of 5′-deleted promoter plasmids were constructed to define the core promoter of <i>CCKAR</i>. The effects of sequence variants located in promoter (PSNP) and conserved (CSNP) regions on promoter activity were analyzed by comparing luciferase activity between haplotypes. A total of 182 variants were found in the 21 kb region. There were no large structural variants around <i>CCKAR</i>. pNL−328/+183, the one with the shortest insertion, showed the highest activity among the six promoter constructs, implying that the key cis elements regulating <i>CCKAR</i> expression are mainly distributed 328 bp upstream. We detected significant activity differences between high- and low-growth associated haplotypes in four of the six promoter constructs. The high-growth haplotypes of constructs pNL−1646/+183, pNL−799/+183 and pNL−528/+183 showed lower activities than the low-growth haplotypes, which is consistent with decreased expression of <i>CCKAR</i> in high-growth chickens. Lower expression of the high-growth allele was also detected for the CSNP5-containing construct. The data suggest that the core promoter of <i>CCKAR</i> is located the 328 bp region upstream from the transcription start site. Lower expression activities shown by the high-growth haplotypes in the reporter assay suggest that CSNP5 and variants located between 328 bp and 1646 bp upstream form a promising molecular basis for decreased expression of <i>CCKAR</i> and increased growth in chickens.https://www.mdpi.com/2073-4425/13/6/1083chicken<i>CCKAR</i>satietygrowthcore promoterexpression activity |
| spellingShingle | Zhepeng Wang Angus M. A. Reid Peter W. Wilson Ian C. Dunn Identification of the Core Promoter and Variants Regulating Chicken <i>CCKAR</i> Expression Genes chicken <i>CCKAR</i> satiety growth core promoter expression activity |
| title | Identification of the Core Promoter and Variants Regulating Chicken <i>CCKAR</i> Expression |
| title_full | Identification of the Core Promoter and Variants Regulating Chicken <i>CCKAR</i> Expression |
| title_fullStr | Identification of the Core Promoter and Variants Regulating Chicken <i>CCKAR</i> Expression |
| title_full_unstemmed | Identification of the Core Promoter and Variants Regulating Chicken <i>CCKAR</i> Expression |
| title_short | Identification of the Core Promoter and Variants Regulating Chicken <i>CCKAR</i> Expression |
| title_sort | identification of the core promoter and variants regulating chicken i cckar i expression |
| topic | chicken <i>CCKAR</i> satiety growth core promoter expression activity |
| url | https://www.mdpi.com/2073-4425/13/6/1083 |
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