Mutations in U6 snRNA that alter splice site specificity: implications for the active site

Science ◽  
1993 ◽  
Vol 262 (5142) ◽  
pp. 1982-1988 ◽  
Author(s):  
C. Lesser ◽  
C Guthrie
Keyword(s):  
Splice Site ◽  
Active Site ◽  
Site Specificity ◽  
U6 Snrna

RNA Splicing by the Spliceosome

2020 ◽  
Vol 89 (1) ◽  
pp. 359-388 ◽  
Author(s):  
Max E. Wilkinson ◽  
Clément Charenton ◽  
Kiyoshi Nagai
Keyword(s):  
Splice Site ◽  
Active Site ◽  
Rna Splicing ◽  
Messenger Rna ◽  
Structural Studies ◽  
U2 Snrna ◽  
U6 Snrna ◽  
Mature Mrna ◽  
Catalytic Metal ◽  
Small Nuclear Ribonucleoproteins

The spliceosome removes introns from messenger RNA precursors (pre-mRNA). Decades of biochemistry and genetics combined with recent structural studies of the spliceosome have produced a detailed view of the mechanism of splicing. In this review, we aim to make this mechanism understandable and provide several videos of the spliceosome in action to illustrate the intricate choreography of splicing. The U1 and U2 small nuclear ribonucleoproteins (snRNPs) mark an intron and recruit the U4/U6.U5 tri-snRNP. Transfer of the 5′ splice site (5′SS) from U1 to U6 snRNA triggers unwinding of U6 snRNA from U4 snRNA. U6 folds with U2 snRNA into an RNA-based active site that positions the 5′SS at two catalytic metal ions. The branch point (BP) adenosine attacks the 5′SS, producing a free 5′ exon. Removal of the BP adenosine from the active site allows the 3′SS to bind, so that the 5′ exon attacks the 3′SS to produce mature mRNA and an excised lariat intron.

scholarly journals Active site specificity profiling datasets of matrix metalloproteinases (MMPs) 1, 2, 3, 7, 8, 9, 12, 13 and 14

Data in Brief ◽  
2016 ◽  
Vol 7 ◽  
pp. 299-310 ◽  
Author(s):  
Ulrich Eckhard ◽  
Pitter F. Huesgen ◽  
Oliver Schilling ◽  
Caroline L. Bellac ◽  
Georgina S. Butler ◽  
...  
Keyword(s):  
Matrix Metalloproteinases ◽  
Active Site ◽  
Site Specificity

X-ray structures of five renin inhibitors bound to saccharopepsin: exploration of active-site specificity 1 1Edited by R. Huber

2000 ◽  
Vol 303 (5) ◽  
pp. 745-760 ◽  
Author(s):  
Nora B Cronin ◽  
Mohammed O Badasso ◽  
Ian J Tickle ◽  
Thomas Dreyer ◽  
Dennis J Hoover ◽  
...  
Keyword(s):  
Active Site ◽  
Site Specificity ◽  
X Ray ◽  
Renin Inhibitors

A New Covalent Inhibitor of Class C β-Lactamases Reveals Extended Active Site Specificity

Biochemistry ◽  
2015 ◽  
Vol 54 (50) ◽  
pp. 7375-7384 ◽  
Author(s):  
Ronak Tilvawala ◽  
Michael Cammarata ◽  
S. A. Adediran ◽  
Jennifer S. Brodbelt ◽  
R. F. Pratt
Keyword(s):  
Active Site ◽  
Site Specificity ◽  
Class C ◽  
Covalent Inhibitor

scholarly journals Structural Mapping of the Active Site Specificity Determinants of Human Tissue-type Plasminogen Activator

1997 ◽  
Vol 272 (35) ◽  
pp. 21713-21719 ◽  
Author(s):  
Martin Renatus ◽  
Wolfram Bode ◽  
Robert Huber ◽  
Jörg Stürzebecher ◽  
Dagmar Prasa ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Active Site ◽  
Human Tissue ◽  
Tissue Type ◽  
Site Specificity ◽  
Structural Mapping ◽  
Tissue Type Plasminogen Activator ◽  
Type Plasminogen Activator ◽  
Specificity Determinants

scholarly journals Mechanism of 5ʹ splice site transfer for human spliceosome activation

Science ◽  
2019 ◽  
Vol 364 (6438) ◽  
pp. 362-367 ◽  
Author(s):  
Clément Charenton ◽  
Max E. Wilkinson ◽  
Kiyoshi Nagai
Keyword(s):  
Splice Site ◽  
Messenger Rna ◽  
Catalytic Center ◽  
U6 Snrna ◽  
Dead Box ◽  
Cryo Electron Microscopy ◽  
Branch Point Sequence ◽  
U1 Snrnp ◽  
Spliceosome Activation ◽  
Small Nuclear Ribonucleoproteins

The prespliceosome, comprising U1 and U2 small nuclear ribonucleoproteins (snRNPs) bound to the precursor messenger RNA 5ʹ splice site (5ʹSS) and branch point sequence, associates with the U4/U6.U5 tri-snRNP to form the fully assembled precatalytic pre–B spliceosome. Here, we report cryo–electron microscopy structures of the human pre–B complex captured before U1 snRNP dissociation at 3.3-angstrom core resolution and the human tri-snRNP at 2.9-angstrom resolution. U1 snRNP inserts the 5ʹSS–U1 snRNA helix between the two RecA domains of the Prp28 DEAD-box helicase. Adenosine 5ʹ-triphosphate–dependent closure of the Prp28 RecA domains releases the 5ʹSS to pair with the nearby U6 ACAGAGA-box sequence presented as a mobile loop. The structures suggest that formation of the 5ʹSS-ACAGAGA helix triggers remodeling of an intricate protein-RNA network to induce Brr2 helicase relocation to its loading sequence in U4 snRNA, enabling Brr2 to unwind the U4/U6 snRNA duplex to allow U6 snRNA to form the catalytic center of the spliceosome.

Base pairing between the 3' exon and an internal guide sequence increases 3' splice site specificity in the Tetrahymena self-splicing rRNA intron

1990 ◽  
Vol 10 (6) ◽  
pp. 2960-2965
Author(s):  
E R Suh ◽  
R B Waring
Keyword(s):  
Splice Site ◽  
Site Specificity ◽  
Group I Introns ◽  
Base Pairing ◽  
Group I ◽  
Guide Sequence ◽  
Compensatory Mutations ◽  
Self Splicing ◽  
Splice Site Usage ◽  
Selection Of

It has been proposed that recognition of the 3' splice site in many group I introns involves base pairing between the start of the 3' exon and a region of the intron known as the internal guide sequence (R. W. Davies, R. B. Waring, J. Ray, T. A. Brown, and C. Scazzocchio, Nature [London] 300:719-724, 1982). We have examined this hypothesis, using the self-splicing rRNA intron from Tetrahymena thermophila. Mutations in the 3' exon that weaken this proposed pairing increased use of a downstream cryptic 3' splice site. Compensatory mutations in the guide sequence that restore this pairing resulted in even stronger selection of the normal 3' splice site. These changes in 3' splice site usage were more pronounced in the background of a mutation (414A) which resulted in an adenine instead of a guanine being the last base of the intron. These results show that the proposed pairing (P10) plays an important role in ensuring that cryptic 3' splice sites are selected against. Surprisingly, the 414A mutation alone did not result in activation of the cryptic 3' splice site.

scholarly journals U1 snRNP-Dependent Function of TIAR in the Regulation of Alternative RNA Processing of the Human Calcitonin/CGRP Pre-mRNA

2003 ◽  
Vol 23 (17) ◽  
pp. 5959-5971 ◽  
Author(s):  
Hui Zhu ◽  
Robert A. Hasman ◽  
Katherine M. Young ◽  
Nancy L. Kedersha ◽  
Hua Lou
Keyword(s):  
Splice Site ◽  
Rna Processing ◽  
Rna Binding ◽  
Sequence Motif ◽  
Human Calcitonin ◽  
Sequence Motifs ◽  
Multiple Sequence ◽  
Enhancer Element ◽  
Binding Domains ◽  
U6 Snrna

ABSTRACT Alternative RNA processing of human calcitonin/CGRP pre-mRNA is regulated by an intronic enhancer element. Previous studies have demonstrated that multiple sequence motifs within the enhancer and a number of trans-acting factors play critical roles in the regulation. Here, we report the identification of TIAR as a novel player in the regulation of human calcitonin/CGRP alternative RNA processing. TIAR binds to the U tract sequence motif downstream of a pseudo 5′ splice site within the previously characterized intron enhancer element. Binding of TIAR promotes inclusion of the alternative 3′-terminal exon located more than 200 nucleotides upstream from the U tract. In cells that preferentially include this exon, overexpression of a mutant TIAR that lacks the RNA binding domains suppressed inclusion of this exon. In this report, we also demonstrate an unusual novel interaction between U6 snRNA and the pseudo 5′ splice site, which was shown previously to bind U1 snRNA. Interestingly, TIAR binding to the U tract sequence depends on the interaction of not only U1 but also U6 snRNA with the pseudo 5′ splice site. Conversely, TIAR binding promotes U6 snRNA binding to its target. The synergistic relationship between TIAR and U6 snRNA strongly suggests a novel role of U6 snRNP in regulated alternative RNA processing.

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