DOI: 10.1261/rna.7186504. Modified nucleotides at the 5′ end of human U2 snRNA are required for spliceosomal E-complex formation

Modified nucleotides at the 5′ end of human U2 snRNA are required for spliceosomal E-complex formation

10.1261/rna.7186504

Crossref journal-article

Cold Spring Harbor Laboratory

RNA (246)

Abstract

U2 snRNA, a key player in nuclear pre-mRNA splicing, contains a 5′-terminal m3G cap and many internal modifications. The latter were shown in vertebrates to be generally required for U2 function in splicing, but precisely which residues are essential and their role in snRNP and/or spliceosome assembly is presently not clear. Here, we investigated the roles of individual modified nucleotides of HeLa U2 snRNA in pre-mRNA splicing, using a two-step in vitro reconstitution/complementation assay. We show that the three pseudouridines and five 2′O-methyl groups within the first 20 nucleotides of U2 snRNA, but not the m3G cap, are required for efficient pre-mRNA splicing. Individual pseudouridines were not essential, but had cumulative effects on U2 function. In contrast, four of five 2′O-methylations (at positions 1, 2, 12, and 19) were individually required for splicing. The in vitro assembly of 17S U2 snRNPs was not dependent on the presence of modified U2 residues. However, individual internal modifications were required for the formation of the ATP-independent early spliceosomal E complex. Our data strongly suggest that modifications within the first 20 nucleotides of U2 play an important role in facilitating the interaction of U2 with U1 snRNP and/or other factors within the E complex.

Bibliography

DÃNMEZ, G., HARTMUTH, K., & LÃHRMANN, R. (2004). Modified nucleotides at the 5â² end of human U2 snRNA are required for spliceosomal E-complex formation. RNA, 10(12), 1925â1933.

Authors 3

GIZEM DÖNMEZ (first)
KLAUS HARTMUTH (additional)
REINHARD LÜHRMANN (additional)

References 42 Referenced 125

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Dates

Type	When
Created	20 years, 9 months ago (Nov. 3, 2004, 3:24 p.m.)
Deposited	1 year, 7 months ago (Jan. 14, 2024, 1:40 p.m.)
Indexed	1 month, 1 week ago (July 20, 2025, 6:47 p.m.)
Issued	20 years, 9 months ago (Nov. 3, 2004)
Published	20 years, 9 months ago (Nov. 3, 2004)
Published Online	20 years, 9 months ago (Nov. 3, 2004)
Published Print	20 years, 8 months ago (Dec. 1, 2004)

Funders 0

None

BibTeX

@article{D_NMEZ_2004, title={Modified nucleotides at the 5′ end of human U2 snRNA are required for spliceosomal E-complex formation}, volume={10}, ISSN={1469-9001}, url={http://dx.doi.org/10.1261/rna.7186504}, DOI={10.1261/rna.7186504}, number={12}, journal={RNA}, publisher={Cold Spring Harbor Laboratory}, author={DÖNMEZ, GIZEM and HARTMUTH, KLAUS and LÜHRMANN, REINHARD}, year={2004}, month=nov, pages={1925–1933} }

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