Characterization of an alternative splicing by a NAGNAG splice acceptor site in the porcine KIT gene
- Authors
- Kim, Jae-Hwan; Zhong, Tao; Cho, In-Cheol; Lim, Hyun-Tae; Yoo, Chae-Kyoung; Kim, Byeong-Woo; Lee, Jun-Heon; Jeon, Jin-Tae
- Issue Date
- Apr-2011
- Publisher
- SPRINGER
- Keywords
- KIT gene; Aalternative splicing; Pig; NAGNAG; Minigene
- Citation
- GENES & GENOMICS, v.33, no.2, pp.179 - 186
- Indexed
- SCIE
SCOPUS
KCI
- Journal Title
- GENES & GENOMICS
- Volume
- 33
- Number
- 2
- Start Page
- 179
- End Page
- 186
- URI
- https://scholarworks.bwise.kr/gnu/handle/sw.gnu/23777
- DOI
- 10.1007/s13258-010-0156-9
- ISSN
- 1976-9571
- Abstract
- The KIT gene has been shown to have multiple functions in hematopoiesis, melanogenesis, and gametogenesis. In addition, mutations of this gene cause pigmentation disorders in humans and mice and are responsible for coat color differences in pigs. While characterizing polymorphisms in the porcine KIT gene, we detected alternative splicing (AS) of the NAGNAG splice acceptor site at the boundary of intron 4 and exon 5. This AS event generated the E and I isoforms, characterized by insertion or deletion, respectively, of CAG at the borders of coding sequence. AS patterns measured in tissue samples from two randomly selected animals did not identified any tissue-specific outcomes. Analysis of AS patterns using three breeds demonstrated that Landrace and Large White pigs expressed both the E and I isoforms. In contrast, a subset of specimens from Korean Native Pigs (KNP) yielded a single I isoform. Alignment of the sequence from several species revealed that the region between the branch point sequence (BPS) and 3' acceptor site is conserved. However, it is appeared that the selection of either the proximal or distal splice site varied between species. To test the breed specificity the NAGNAG splice acceptor site, we constructed two lineages of minigenes from KNP and Landrace pigs harboring breed-specific mutations. The minigene splicing assay demonstrated that both types of minigenes expressed both the E and I isoforms in two host cell lines, and no differences were detected in the AS pattern between the two breeds. We conclude that the AS at the NAGNAG splice acceptor site on intron 4/exon 5 in the porcine KIT gene is the result of noise selection at the splice site by the splicing machinery. Therefore, this AS event in the porcine KIT gene is unlikely to have any relationship with the coat color variations of Landrace and KNP breeds.
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