期刊
G3-GENES GENOMES GENETICS
卷 8, 期 4, 页码 1367-1377出版社
GENETICS SOCIETY AMERICA
DOI: 10.1534/g3.118.200022
关键词
alternative splicing; Arabidopsis thaliana; PRP18; second step factor
资金
- Institute of Plant and Microbial Biology (IPMB) at Academia Sinica
- Taiwan Ministry of Science and Technology [MOST 105-2311-B-001-071, MOST 106-2311-B-001-023-MY2, MOST 103-2311-B001-004-MY3]
- Academia Sinica
Splicing of pre-mRNA involves two consecutive trans-esterification steps that take place in the spliceosome, a large dynamic ribonucleoprotein complex situated in the nucleus. In addition to core spliceosomal proteins, each catalytic step requires step-specific factors. Although the Arabidopsis thaliana genome encodes around 430 predicted splicing factors, functional information about these proteins is limited. In a forward genetic screen based on an alternatively-spliced GFP reporter gene in Arabidopsis thaliana, we identified a mutant impaired in putative step II factor PRP18a, which has not yet been investigated for its role in pre-mRNA splicing in plants. Step II entails cleavage at the 3 splice site accompanied by ligation of the 5 and 3 exons and intron removal. In the prp18 mutant, splicing of a U2-type intron with non-canonical AT-AC splice sites in GFP pre-mRNA is reduced while splicing of a canonical GT-AG intron is enhanced, resulting in decreased levels of translatable GFP mRNA and GFP protein. These findings suggest that wild-type PRP18a may in some cases promote splicing at weak, non-canonical splice sites. Analysis of genome-wide changes in alternative splicing in the prp18a mutant identified numerous cases of intron retention and a preponderance of altered 3 splice sites, suggesting an influence of PRP18a on 3 splice site selection. The prp18a mutant featured short roots on synthetic medium and small siliques, illustrating that wild-type PRP18a function is needed for a normal phenotype. Our study expands knowledge of plant splicing factors and provides foundational information and resources for further functional studies of PRP18 proteins in plants.
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