Metal binding and substrate positioning by evolutionarily invariant U6 sequences in catalytically active protein-free snRNAs.
We have previously shown that a base-paired complex formed by two of the spliceosomal RNA components, U6 and U2 small nuclear RNAs (snRNAs), can catalyze a two-step splicing reaction that depended on an evolutionarily invariant region in U6, the ACAGAGA box. Here we further analyze this RNA-catalyzed reaction and show that while the 5' and 3' splice site substrates are juxtaposed and positioned near the ACAGAGA sequence in U6, the role of the snRNAs in the reaction is beyond mere juxtaposition of the substrates and likely involves the formation of a sophisticated active site. Interestingly, the snRNA-catalyzed reaction is metal dependent, as is the case with other known splicing RNA enzymes, and terbium(III) cleavage reactions indicate metal binding by the U6/U2 complex within the evolutionarily conserved regions of U6. The above results, combined with the structural similarities between U6 and catalytically critical domains in group II self-splicing introns, suggest that the base-paired complex of U6 and U2 snRNAs is a vestigial ribozyme and a likely descendant of a group II-like self-splicing intron.
Lee
Caroline
C
Center for RNA Molecular Biology, Case Western Reserve University, Cleveland, Ohio 44106, USA.
Jaladat
Yasaman
Y
Mohammadi
Afshin
A
Sharifi
Armin
A
Geisler
Sarah
S
Valadkhan
Saba
S
D001483:Base Sequence; D005091:Exons; D006801:Humans; D008274:Magnesium / Q000378:metabolism*; D008969:Molecular Sequence Data; D012326:RNA Splicing; D016337:RNA, Catalytic / Q000378:metabolism*; D012342:RNA, Small Nuclear / Q000378:metabolism*; D013379:Substrate Specificity
D016337:RNA, Catalytic; D012342:RNA, Small Nuclear; C405484:U2 small nuclear RNA; C405486:U6 small nuclear RNA; D008274:Magnesium
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