Effects of splice sites on the intron retention in histamine H3 receptors from rats and mice.
In the alternative splicing, intron retention, of histamine H(3) receptors in rats and mice, the short transcript isoforms that are excised alternatively spliced introns are easily detected in a very low level in rats and are undetectable in mice using the regular PCR protocol. The retained introns have common 5' splice site and different 3' splice sites. The detailed mechanism for the special alternative splicing remains largely unclear. In this study, we developed a minigene splicing system to recapitulate natural alternative splicing of the receptors and investigated the effects of 5' and 3' splice sites on intron retention in HeLa cells. Mutating weak 5' and 3' splice sites of the alternatively spliced introns toward the canonical consensus sequences promoted the splicing of the corresponding introns in rat and mouse minigenes. The effect of splice site strength was context-dependent and much more significant for the 3' splice site of the longer alternative intron than for the 3' splice site of the shorter alternative intron and the common 5' splice sites; it was also more significant in the rat minigene than in the mouse minigene. Mutating the 3' splice site of the longer alternative intron resulted in almost complete splicing of the intron and made the corresponding isoform to become the nearly exclusive transcript in the rat minigene.
Ding
Wenyong
W
Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian 116044, China.
Lin
Lin
L
Ren
Feng
F
Zou
Hanfa
H
Duan
Ziyuan
Z
Dai
Jianwu
J
D017398:Alternative Splicing; D000818:Animals; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D051379:Mice; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D009154:Mutation; D022821:RNA Splice Sites; D051381:Rats; D018100:Receptors, Histamine H3 / Q000235:genetics*; D016415:Sequence Alignment
D022821:RNA Splice Sites; D018100:Receptors, Histamine H3