Splicing proofreading at 5′ splice sites by ATPase Prp28p

Fei Yang, Xiu Ye Wang, Zhi Min Zhang, Jia Pu, Yu Jie Fan, Jiahai Zhou, Charles C. Query, Yong Zhen Xu

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Fidelity and efficiency of pre-mRNA splicing are critical for generating functional mRNAs, but how such accuracy in 5′ splice site (SS) selection is attained is not fully clear. Through a series of yeast genetic screens, we isolated alleles of prp28 that improve splicing of suboptimal 5′SS substrates, demonstrating that WT-Prp28p proofreads, and consequently rejects, poor 5′SS. Prp28p is thought to facilitate the disruption of 5′SS-U1 snRNA pairing to allow for 5′SS-U6 snRNA pairing in the catalytic spliceosome; unexpectedly, 5′SS proofreading by Prp28p is dependent on competition with the stability of the 5′SS:U6 duplex, but not the 5′SS:U1 duplex. E404K, the strongest prp28 allele containing a mutation located in the linker region between adenosine triphosphatase (ATPase) subdomains, exhibited lower RNA-binding activity and enhanced splicing of suboptimal substrates before first-step catalysis, suggesting that decreased Prp28p activity allows longer time for suboptimal 5′SS substrates to pair with U6 snRNA and thereby reduces splicing fidelity. Residue E404 is critical for providing high splicing activity, demonstrated here in both yeast and Drosophila cells. Thus, the subdomain linker in Prp28p plays important roles both in splicing efficiency across species and in proofreading of 5′SS.

Original languageEnglish (US)
Pages (from-to)4660-4670
Number of pages11
JournalNucleic Acids Research
Volume41
Issue number8
DOIs
StatePublished - Apr 2013

Fingerprint

RNA Splice Sites
Adenosine Triphosphatases
Yeasts
Alleles
Spliceosomes
RNA Precursors
Catalysis
Drosophila
RNA
Messenger RNA
Mutation
U6 small nuclear RNA
U1 small nuclear RNA

ASJC Scopus subject areas

  • Genetics

Cite this

Yang, F., Wang, X. Y., Zhang, Z. M., Pu, J., Fan, Y. J., Zhou, J., ... Xu, Y. Z. (2013). Splicing proofreading at 5′ splice sites by ATPase Prp28p. Nucleic Acids Research, 41(8), 4660-4670. https://doi.org/10.1093/nar/gkt149

Splicing proofreading at 5′ splice sites by ATPase Prp28p. / Yang, Fei; Wang, Xiu Ye; Zhang, Zhi Min; Pu, Jia; Fan, Yu Jie; Zhou, Jiahai; Query, Charles C.; Xu, Yong Zhen.

In: Nucleic Acids Research, Vol. 41, No. 8, 04.2013, p. 4660-4670.

Research output: Contribution to journalArticle

Yang, F, Wang, XY, Zhang, ZM, Pu, J, Fan, YJ, Zhou, J, Query, CC & Xu, YZ 2013, 'Splicing proofreading at 5′ splice sites by ATPase Prp28p', Nucleic Acids Research, vol. 41, no. 8, pp. 4660-4670. https://doi.org/10.1093/nar/gkt149
Yang F, Wang XY, Zhang ZM, Pu J, Fan YJ, Zhou J et al. Splicing proofreading at 5′ splice sites by ATPase Prp28p. Nucleic Acids Research. 2013 Apr;41(8):4660-4670. https://doi.org/10.1093/nar/gkt149
Yang, Fei ; Wang, Xiu Ye ; Zhang, Zhi Min ; Pu, Jia ; Fan, Yu Jie ; Zhou, Jiahai ; Query, Charles C. ; Xu, Yong Zhen. / Splicing proofreading at 5′ splice sites by ATPase Prp28p. In: Nucleic Acids Research. 2013 ; Vol. 41, No. 8. pp. 4660-4670.
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