Journal
ANIMAL GENETICS
Volume 45, Issue 5, Pages 641-651Publisher
WILEY
DOI: 10.1111/age.12182
Keywords
bovine; functional SNP; katanin p60 subunit A-like 1; promoter luciferase activity analysis; semen traits; splice variant
Funding
- National Science and Technology Pillar Program during the 12th Five-Year Plan of China [2011BAD19B04]
- Modern Agro-industry Technology Research System [CARS-37]
- State Major Project of Transgenic [2011ZX08007-004]
- Well-bred Program of Shandong Province [2013LZ10-06]
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Katanin p60 subunit A-like 1 (KATNAL1) is an ATPase that regulates Sertoli cell microtubule dynamics and sperm retention. We evaluated one novel splice variant and characterized the promoter and a functional single nucleotide polymorphism (SNP) of the bovine KATNAL1 gene to explore its expression pattern, possible regulatory mechanism and relationship with semen traits in Chinese Holstein bulls. A novel splice variant, KATNAL1 transcript variant 2 (KATNAL1-TV2) of the retained 68bp in intron 2, was identified by RT-PCR and compared with KATNAL1 transcript variant 1 (KATNAL1-TV1, NM 001192918.1) in various tissues. Bioinformatics analyses predicted that KATNAL1 transcription was regulated by two promoters: P1 in KATNAL1-TV1 and P2 in KATNAL1-TV2. Results of qRT-PCR revealed that KATNAL1-TV1 had higher expression than did KATNAL1-TV2 in testes of adult bulls (P<0.05). Promoter luciferase activity analysis suggested that the core sequences of P1 and P2 were mapped to the region of c.-575c.-180 and c.163-40c.333+59 respectively. One novel SNP (c.163-210T>C, ss836312085) located in intron 1 was found using sequence alignment. The SNP in P2 resulted in the presence of the DeltaE binding site, improving its basal promoter activity (P<0.05); and we observed a greater sperm deformity rate in bulls with the genotype CC than in those with the genotype TT (P<0.05), which indicated that different genotypes were associated with the bovine semen traits. Bioinformatics analysis of the KATNAL1 protein sequence predicted that the loss of the MIT domain in the KATNAL1-TV2 transcript resulted in protein dysfunction. These findings help us to understand that a functional SNP in P2 and subsequent triggering of expression diversity of KATNAL1 transcripts are likely to play an important role with regard to semen traits in bull breeding programs.
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