The U1 small nuclear RNA-protein complex selectively binds a 5′ splice site in vitro

TitleThe U1 small nuclear RNA-protein complex selectively binds a 5′ splice site in vitro
Publication TypeJournal Articles
Year of Publication1983
AuthorsMount SM, Pettersson I, Hinterberger M, Karmas A, Steitz JA
JournalCell
Volume33
Issue2
Pagination509 - 518
Date Published1983/06//
ISBN Number00928674
Abstract

The ability of purified U1 small nuclear RNA-protein complexes (U1 snRNPs) to bind in vitro to two RNAs transcribed from recombinant DNA clones by bacteriophage T7 RNA polymerase has been studied. A transcript which contains sequences corresponding to the small intron and flanking exons of the major mouse beta-globin gene is bound in marked preference to an RNA devoid of splice site sequences. The site of U1 snRNP binding to the globin RNA has been defined by T1 ribonuclease digestion of the RNA-U1 snRNP complex. A 15-17-nucleotide region, including the 5' splice site, remains undigested and complexed with the snRNP such that it can be co-precipitated by antibodies directed against the U1 snRNP. Partial proteinase K digestion of the U1 snRNP abolishes interaction with the globin RNA, indicating that the snRNP proteins contribute significantly to RNA binding. No RNA cleavage, splicing, or recognition of the 3' splice site by U1 snRNPs has been detected. Our results are discussed in terms of the probable role of U1 snRNPs in the messenger RNA splicing of eucaryotic cell nuclei.

URLhttp://ukpmc.ac.uk/abstract/MED/6190573
DOI10.1016/0092-8674(83)90432-4