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Biochemical Genetics

, Volume 57, Issue 6, pp 801–812 | Cite as

Tissue-Specific Monoallelic Expression of Bovine AXL is Associated with DNA Methylation of Promoter DMR

  • Da Xu
  • Guannan Wang
  • Junliang Li
  • Cui Zhang
  • Weina Chen
  • Dongjie Li
  • Shijie LiEmail author
Original Article

Abstract

The AXL protein is a receptor tyrosine kinase and is often implicated in proliferation, migration and therapy resistance in various cancers. The AXL gene in humans is maternally expressed and paternally imprinted with differentially methylated regions (DMR) surrounding the promoter region. However, the imprinting status and epigenetic regulation of AXL gene in cattle remain unclear. Therefore, we explored the molecular structure along with the patterns of allelic expression and DNA methylation of the bovine AXL gene. First, the complete cDNA sequence of bovine AXL was gathered by Sanger method, from transcripts obtained from RT-PCR, 5′ and 3′ -RACE. In silico BLAST alignments showed that the longest mRNA sequence of bovine AXL consists of 19 exons and encodes a protein of 887 amino acids. We further analyzed the allelic expression of bovine AXL by employing single-nucleotide polymorphism (SNP)-based sequencing method. A SNP site (GenBank Accession no: rs210020651) found in exon 7 allowed us to distinguish the two parental alleles. Monoallelic expression of AXL was observed in four adult bovine tissues (heart, liver, spleen and fat), while biallelic expression was found in the other adult tissues such as the lung, kidney, muscle, brain and placenta. To determine whether the DNA methylation played a role in the tissue-specific imprinting of bovine AXL, we performed bisulfite sequencing of two regions: region 1 was a CpG island (CGI) in AXL promoter, mapping to 643 bp upstream of the transcription start site of AXL 5′-v1 transcripts, while region two was homologous to the region of human AXL DMR, with 10 CpG sites overlapping the first translation start site (TSS1) of bovine AXL. In region 2, DNA from both monoallelic and biallelic expressed tissues were mostly found to be completely unmethylated. However, tissue-specific differential methylation patterns were found in monoallelic expressed tissues such as the heart and liver while hypomethylation was noted in the promoter CpG island in biallelic expressed tissues such as the lung. These observations demonstrated that the tissue-specific monoallelic expression of bovine AXL is dependent on the DNA methylation of its promoter region.

Keywords

Tissue-specific DMR AXL Bovine 

Notes

Acknowledgements

This study was funded by Hebei province Natural Science Foundation of China (Grant No. C2016204092) and The National Natural Science Foundation of China (Grant No. 31372312).

Compliance with Ethical Standards

Conflicts of interest

The authors declare that they have no conflict of interest.

Informed Consent

Informed consent was obtained from the owner of the animal. All institutional and national guidelines for the care and use of laboratory animals were followed, and all animal experiments were approved by the Agriculture Research Animal Care Committee of Hebei Agriculture University.

Supplementary material

10528_2019_9925_MOESM1_ESM.docx (15 kb)
Supplementary material 1 (DOCX 14 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.College of Life ScienceAgricultural University of HebeiBaodingChina
  2. 2.College of Life ScienceCangzhou Normal UniversityCangzhouChina
  3. 3.Department of Traditional Chinese MedicineHebei UniversityBaodingChina
  4. 4.College of Bioscience and BioengineeringHebei University of Science and TechnologyShijiazhuangChina

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