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Genetic interactions between the 5' and 3' splice site consensus sequences and U6 snRNA during the second catalytic step of pre-mRNA splicing.
RNA. 2001 Dec; 7(12):1845-54.RNA

Abstract

The YAG/ consensus sequence at the 3' end of introns (the slash indicates the location of the 3' splice site) is essential for catalysis of the second step of pre-mRNA splicing. Little is known about the interactions formed by these three nucleotides in the spliceosome. Although previous observations have suggested that the G of the YAG/ interacts with the first nucleotide of the /GUA consensus sequence at the 5' end of the intron, additional interactions have not been identified. Here we report several striking genetic interactions between A+3 of the 5' /GUA with Y-3 of the 3' YAG/ and G50 of the highly conserved ACAGAG motif in U6 snRNA. Two mutations in U6 G50 of the ACAGAG can weakly suppress two mutations in A+3 of the 5' /GUA. This suppression is significantly enhanced upon the inclusion of a specific mutation Y-3 in the 3' YAG/. RNA analysis confirmed that the severe splicing defect observed in A+3 and Y-3 double mutants can be rescued to near wild-type levels by the mutations in U6 G50. The contributions of each mutation to the genetic interaction and the strong position specificity of suppression, combined with previous findings, support a model in which the 5' /GUA and the GAG of U6 function in binding the 3' YAG/ during the second catalytic step.

Authors+Show Affiliations

Department of Biochemistry and Biophysics, University of California, San Francisco 94143, USA.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

11780639

Citation

Collins, C A., and C Guthrie. "Genetic Interactions Between the 5' and 3' Splice Site Consensus Sequences and U6 snRNA During the Second Catalytic Step of pre-mRNA Splicing." RNA (New York, N.Y.), vol. 7, no. 12, 2001, pp. 1845-54.
Collins CA, Guthrie C. Genetic interactions between the 5' and 3' splice site consensus sequences and U6 snRNA during the second catalytic step of pre-mRNA splicing. RNA. 2001;7(12):1845-54.
Collins, C. A., & Guthrie, C. (2001). Genetic interactions between the 5' and 3' splice site consensus sequences and U6 snRNA during the second catalytic step of pre-mRNA splicing. RNA (New York, N.Y.), 7(12), 1845-54.
Collins CA, Guthrie C. Genetic Interactions Between the 5' and 3' Splice Site Consensus Sequences and U6 snRNA During the Second Catalytic Step of pre-mRNA Splicing. RNA. 2001;7(12):1845-54. PubMed PMID: 11780639.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Genetic interactions between the 5' and 3' splice site consensus sequences and U6 snRNA during the second catalytic step of pre-mRNA splicing. AU - Collins,C A, AU - Guthrie,C, PY - 2002/1/10/pubmed PY - 2002/1/29/medline PY - 2002/1/10/entrez SP - 1845 EP - 54 JF - RNA (New York, N.Y.) JO - RNA VL - 7 IS - 12 N2 - The YAG/ consensus sequence at the 3' end of introns (the slash indicates the location of the 3' splice site) is essential for catalysis of the second step of pre-mRNA splicing. Little is known about the interactions formed by these three nucleotides in the spliceosome. Although previous observations have suggested that the G of the YAG/ interacts with the first nucleotide of the /GUA consensus sequence at the 5' end of the intron, additional interactions have not been identified. Here we report several striking genetic interactions between A+3 of the 5' /GUA with Y-3 of the 3' YAG/ and G50 of the highly conserved ACAGAG motif in U6 snRNA. Two mutations in U6 G50 of the ACAGAG can weakly suppress two mutations in A+3 of the 5' /GUA. This suppression is significantly enhanced upon the inclusion of a specific mutation Y-3 in the 3' YAG/. RNA analysis confirmed that the severe splicing defect observed in A+3 and Y-3 double mutants can be rescued to near wild-type levels by the mutations in U6 G50. The contributions of each mutation to the genetic interaction and the strong position specificity of suppression, combined with previous findings, support a model in which the 5' /GUA and the GAG of U6 function in binding the 3' YAG/ during the second catalytic step. SN - 1355-8382 UR - https://www.unboundmedicine.com/medline/citation/11780639/Genetic_interactions_between_the_5'_and_3'_splice_site_consensus_sequences_and_U6_snRNA_during_the_second_catalytic_step_of_pre_mRNA_splicing_ L2 - http://www.rnajournal.org/cgi/pmidlookup?view=long&pmid=11780639 DB - PRIME DP - Unbound Medicine ER -