title: Association between an alternative promoter polymorphism and sperm deformity rate is due to modulation of the expression of KATNAL1 transcripts in Chinese Holstein bulls.
abstract: 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 68 bp 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.-575˜c.-180 and c.163-40˜c.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.
authors.affiliation: College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China; Dairy Cattle Research Center, Shandong Academy of Agricultural Science, Jinan, 250131, China; College of Animal Science, Henan Institute of Science and Technology, Xinxiang, 453000, China.
authors.forename: X
authors.initials: X
authors.lastname: Zhang
authors.forename: C
authors.initials: C
authors.lastname: Wang
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: Z
authors.initials: Z
authors.lastname: Ju
authors.forename: C
authors.initials: C
authors.lastname: Qi
authors.forename: X
authors.initials: X
authors.lastname: Wang
authors.forename: J
authors.initials: J
authors.lastname: Huang
authors.forename: S
authors.initials: S
authors.lastname: Zhang
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.forename: J
authors.initials: J
authors.lastname: Zhong
authors.forename: F
authors.initials: F
authors.lastname: Shi
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics*; D000818:Animals; D002417:Cattle / Q000235:genetics*; D019295:Computational Biology; D005838:Genotype; D008297:Male; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic; D020033:Protein Isoforms / Q000235:genetics; D012661:Semen / Q000502:physiology; D055101:Semen Analysis; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013094:Spermatozoa / Q000473:pathology*; D013737:Testis / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D020033:Protein Isoforms; D000251:Adenosine Triphosphatases
keywords: bovine; functional SNP; katanin p60 subunit A-like 1; promoter luciferase activity analysis; semen traits; splice variant