Domain analysis of human U5 RNA - Cap trimethylation, protein binding, and spliceosome assembly - Research portal of Justus Liebig University Giessen:

Journal article

Domain analysis of human U5 RNA - Cap trimethylation, protein binding, and spliceosome assembly

Authors list: Hinz, M; Moore, MJ; Bindereif, A

Publication year: 1996

Pages: 19001-19007

Journal: Journal of Biological Chemistry

Volume number: 271

Issue number: 31

ISSN: 0021-9258

eISSN: 1083-351X

DOI Link: https://doi.org/10.1074/jbc.271.31.19001

Publisher: Elsevier

Abstract:
We have analyzed the sequence requirements of the human U5 RNA during small nuclear ribonucleoprotein (snRNP) and spliceosome assembly. A collection of mutant derivatives of the human U5 RNA gene was constructed in a U1 expression vector and transiently transfected in mammalian cells. Using immunoprecipitation and affinity selection assays, the cap trimethylation, the binding of Sm proteins and of the U5 snRNP-specific protein p220, as well as the assembly of the U4/U5/U6 triple snRNP and of spliceosomes were determined. By mutational analysis we were able to assign distinct functions to several structural elements of the human U5 RNA. Efficient binding of the Sm proteins requires the 3' stem-loop. Both the Sm protein-binding site and the 3' stem-loop are necessary for the formation of the trimethyl guanosine cap, consistent with Sm protein binding being a prerequisite for cap trimethylation. Specific elements of the U5 RNA 5' stem-loop contribute to efficient p220 association, in particular stem Ib. Interestingly, the highly conserved loop I appears to be a multifunctional element; in addition to its function in splice-site selection the 5' loop is involved in binding of p220 and in the assembly of the U4/U5/U6 triple snRNP. In sum, this mutational analysis has identified four functional domains of the human U5 RNA.

Citation Styles

Harvard Citation style: Hinz, M., Moore, M. and Bindereif, A. (1996) Domain analysis of human U5 RNA - Cap trimethylation, protein binding, and spliceosome assembly, Journal of Biological Chemistry, 271(31), pp. 19001-19007. https://doi.org/10.1074/jbc.271.31.19001

APA Citation style: Hinz, M., Moore, M., & Bindereif, A. (1996). Domain analysis of human U5 RNA - Cap trimethylation, protein binding, and spliceosome assembly. Journal of Biological Chemistry. 271(31), 19001-19007. https://doi.org/10.1074/jbc.271.31.19001