Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2007 Dec 15;36(3):814–825. doi: 10.1093/nar/gkm1098

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2007 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

PMC Copyright notice

Figure 4. — The U2-pre-mRNA crosslink occurs in prespliceosomes. (A) The U2-pre-mRNA crosslink at (−6) occurs before tri-snRNP addition to the spliceosome. Extracts were depleted of U6 by pre-treating with an oligonucleotide complementary to U6 (lane 2) or water (lane 1) before the addition of CYH2 pre-mRNA with 4-thioU at (−6), followed by splicing for 20 min then UV irradiation. Isolated RNA was analyzed by 4% denaturing PAGE. The pre-mRNA and four major crosslinked species are indicated on the right. Markers (lane 1), ³²P-labeled pBR322 Msp1-digested DNA. (B) A 2′-O-Me RNA oligonucleotide complementary to the CYH2 branchpoint inhibits the first step of splicing. Body labeled CYH2 was hybridized in splicing buffer with 0, 0.3, 0.5 or 0.7 μg 2′ O-Me RNA oligonucleotide complementary to the branchpoint (lanes 2–5). Splicing was carried out for 20 min and isolated RNA was analyzed by denaturing PAGE. The splicing intermediates and products indicated on the right. Markers (lane 1), as in (A). (C) The U2-pre-mRNA crosslink at (−6) requires prior U2-branchpoint interaction. CYH2 pre-mRNA with 4-thioU at position (−6) was hybridized to 0.5 μg of 2′-O-Me RNA oligonucleotide complementary to the branchpoint (lane 3) or water (lane 2). Extract was added and splicing carried out for 20 min before UV irradiation. Isolated RNA was analyzed by 4% denaturing PAGE. The pre-mRNA and four major crosslinked species are indicated on the right. Markers (lane 1), as in (A).