Molecular Characterization and Phylogenetic Analysis of Casein Gene Family in Camelus ferus

Parveen, Shakeela; Zhu, Peng; Shafique, Laiba; Lan, Hong; Xu, Dingyun; Ashraf, Sana; Ashraf, Saba; Sherazi, Maryam; Liu, Qingyou

doi:10.3390/genes14020256

Cited by 3 publications

(4 citation statements)

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“…Only a few SNPs (4 out of 18 in Bactrian and 2 out of 16 in dromedary) were found in the retrotransposons, contributing to 17.6% of the total genetic diversity found. This result reflects a lower level of genetic diversity found in the casein genes of camelids [3,14,[18][19][20]22,23,61,62] compared to ruminants [13,[63][64][65][66]. These findings are consistent with the observations made by other authors, indicating a significant reduction in genetic diversity due to at least two bottlenecks during the history of camel domestication [56,67,68].…”

Section: Discussionsupporting

confidence: 91%

Sequencing and Characterization of αs2-Casein Gene (CSN1S2) in the Old-World Camels Have Proven Genetic Variations Useful for the Understanding of Species Diversification

Pauciullo,

Versace,

Gaspa

et al. 2023

Animals

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The CSN1S2 gene encodes αs2-casein, the third most abundant protein in camel milk. Despite its importance in foals, human nutrition, and dairy processing, the CSN1S2 gene in camels has received little attention. This study presents the first complete characterization of the CSN1S2 gene sequence in Old-World camels (Camelus bactrianus and Camelus dromedarius). Additionally, the gene promoter, consisting of 752 bp upstream of exon 1, was analyzed. The entire gene comprises 17 exons, ranging in length from 24 bp (exons 4, 8, 11, and 13) to 280 bp (exon 17). Interesting was the identification of the exon 12 in both species. The promoter analysis revealed 24 putative binding sites in the Bactrian camel and 22 in dromedary camel. Most of these sites were typical elements associated with milk protein, such as C/EBP-α, C/EBP-β, Oct-1, and AP1. The SNP discovery showed relatively high genetic diversity compared to other camel casein genes (CSN1S1, CSN2, and CSN3), with a total of 34 polymorphic sites across the two species. Particularly noteworthy is the transition g.311G>A in the CSN1S2 promoter, creating a new putative consensus binding site for a C/EBP-β in the Bactrian camel. At the exon level, two novel variants were found. One was detected in exon 6 of the Bactrian camel (g.3639C>G), resulting in an amino acid replacement, p.36Ile>Met. The second variant was found in noncoding exon 17 of dromedary CSN1S2 (g.1511G>T). Although this mutation occurs in the 3′-UnTranslated Region, it represents the first example of exonic polymorphism in the CSN1S2 for this species. This SNP also affects the binding sites of different microRNAs, including the seed sequence of the miRNA 4662a-3p, highlighting its role as a regulatory factor for CSN1S2 gene. A PCR-RFLP was set up for genotyping a dromedary Tunisian population (n = 157), and the minor allele frequency was found to be 0.27 for the G allele, indicating a potential yield improvement margin. The interspersed elements (INEs) analysis revealed 10 INEs covering 7.34% and 8.14% of the CSN1S2 sequence in the Bactrian and dromedary camels, respectively. Furthermore, six elements (A, B, F, H, I, and L) are shared among cattle and camels and are partially found in other ruminants, suggesting a common ancestral origin of these retrotransposons. Conversely, elements C, D, E, and G are specific to camels.

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Section: Discussionsupporting

confidence: 91%

Sequencing and Characterization of αs2-Casein Gene (CSN1S2) in the Old-World Camels Have Proven Genetic Variations Useful for the Understanding of Species Diversification

Pauciullo,

Versace,

Gaspa

et al. 2023

Animals

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“…Casein is a unique protein in milk . CSN2 is a member of the casein family . We found that almost all isolated cells could express CSN2, indicating that the isolated cells had lactation function.…”

Section: Discussionmentioning

confidence: 73%

“…29 CSN2 is a member of the casein family. 30 We found that almost all isolated cells could express CSN2, indicating that the isolated cells had lactation function. In summary, the cells we isolated could be used for subsequent research.…”

Section: Functional Prediction Of Mir-200amentioning

confidence: 70%

Bta-miR-200a Regulates Milk Fat Biosynthesis by Targeting IRS2 to Inhibit the PI3K/Akt Signal Pathway in Bovine Mammary Epithelial Cells

Tan,

Yang,

Qiu

et al. 2024

J. Agric. Food Chem.

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Milk fat synthesis has garnered significant attention due to its influence on the quality of milk. Recently, an increasing amount of proofs have elucidated that microRNAs (miRNAs) are important post-transcriptional factor involved in regulating gene expression and play a significant role in milk fat synthesis. MiR-200a was differentially expressed in the mammary gland tissue of dairy cows during different lactation periods, which indicated that miR-200a was a candidate miRNA involved in regulating milk fat synthesis. In our research, we investigated the potential function of miR-200a in regulating milk fat biosynthesis in bovine mammary epithelial cells (BMECs). We discovered that miR-200a inhibited cellular triacylglycerol (TAG) synthesis and suppressed lipid droplet formation; at the same time, miR-200a overexpression suppressed the mRNA and protein expression of milk fat metabolismrelated genes, such as fatty acid synthase (FASN), peroxisome proliferator-activated receptor gamma (PPARγ), sterol regulatory element-binding protein 1 (SREBP1), CCAAT enhancer binding protein alpha (CEBPα), etc. However, knocking down miR-200a displayed the opposite results. We uncovered that insulin receptor substrate 2 (IRS2) was a candidate target gene of miR-200a through the bioinformatics online program TargetScan. Subsequently, it was confirmed that miR-200a directly targeted the 3′untranslated region (3′-UTR) of IRS2 via real-time fluorescence quantitative PCR (RT-qPCR), western blot analysis, and dualluciferase reporter gene assay. Additionally, IRS2 knockdown in BMECs has similar effects to miR-200a overexpression. Our research set up the mechanism by which miR-200a interacted with IRS2 and discovered that miR-200a targeted IRS2 and modulated the activity of the PI3K/Akt signaling pathway, thereby taking part in regulating milk fat synthesis in BMECs. Our research results provided valuable information on the molecular mechanisms for enhancing milk quality from the view of miRNA-mRNA regulatory networks.

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“…Few isolated studies have been there for unconventional species. A phylogenetic study of different casein proteins of Camelus ferus depicted distinct clusters of different proteins (Parveen et al, 2023) [6] . Comparative genomic analysis provides a better insight on overall functionality and antimicrobial nature of abundant milk protein casein.…”

Section: Introductionmentioning

confidence: 99%

Comparative genomic analysis for phylogeny, physio-chemical and anti-microbial properties in unconventional dairy animals

Singh,

Chinnareddyvari

et al. 2024

Int. J. Res. Agron.

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Casein is a major milk protein that affects the chemical and anti-microbial properties of milk. Four casein proteins i.e. alphaS1, alphaS2, beta, and kappa-were computed for the construction of a phylogenetic tree. Moreover, computational analysis was also performed for the physio-chemical and anti-microbial properties of milk. Whole protein sequences were retrieved from the NCBI database. Physiochemical properties and anti-microbial peptides were predicted using the ProtParam tool and AMPA Tool, respectively. Phylogenetic analysis revealed strong relatedness between Camel and Yak as compared to Donkey. Moreover, most of the casein proteins were found to be unstable in the instability index. Few bactericidal stretches were found especially alphaS2 casein protein in Camel, Yak and Donkey. The average anti-microbial index was around 0.25 -0.26. This study revealed about phylogenetic relatedness of casein protein in different species. It also highlighted the importance of the bioinformatics approach for better prediction of physio-chemical and anti-microbial features of casein micelles.

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Molecular Characterization and Phylogenetic Analysis of Casein Gene Family in Camelus ferus

Cited by 3 publications

References 59 publications

Sequencing and Characterization of αs2-Casein Gene (CSN1S2) in the Old-World Camels Have Proven Genetic Variations Useful for the Understanding of Species Diversification

Sequencing and Characterization of αs2-Casein Gene (CSN1S2) in the Old-World Camels Have Proven Genetic Variations Useful for the Understanding of Species Diversification

Bta-miR-200a Regulates Milk Fat Biosynthesis by Targeting IRS2 to Inhibit the PI3K/Akt Signal Pathway in Bovine Mammary Epithelial Cells

Comparative genomic analysis for phylogeny, physio-chemical and anti-microbial properties in unconventional dairy animals

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