English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Dynamic contacts of U2, RES, Cwc25, Prp8 and Prp45 proteins with the Pre-mRNA branch-site and 3' splice site during catalytic activation and step 1 catalysis in yeast spliceosomes.

Schneider, C., Agafonov, D. E., Schmitzova, J., Hartmuth, K., Fabrizio, P., & Lührmann, R. (2015). Dynamic contacts of U2, RES, Cwc25, Prp8 and Prp45 proteins with the Pre-mRNA branch-site and 3' splice site during catalytic activation and step 1 catalysis in yeast spliceosomes. PLoS Genetics, 11(9): e1005539. doi:10.1371/journal.pgen.1005539.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-8EF3-B Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002A-22FD-8
Genre: Journal Article

Files

show Files
hide Files
:
2214427.pdf (Publisher version), 7MB
Name:
2214427.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-

Creators

show
hide
 Creators:
Schneider, C.1, Author              
Agafonov, D. E.1, Author              
Schmitzova, J.1, Author              
Hartmuth, K.1, Author              
Fabrizio, P.1, Author              
Lührmann, R.1, Author              
Affiliations:
1Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578576              

Content

show
hide
Free keywords: -
 Abstract: Little is known about contacts in the spliceosome between proteins and intron nucleotides surrounding the pre-mRNA branch-site and their dynamics during splicing. We investigated protein-pre-mRNA interactions by UV-induced crosslinking of purified yeast Bact spliceosomes formed on site-specifically labeled pre-mRNA, and analyzed their changes after conversion to catalytically-activated B* and step 1 C complexes, using a purified splicing system. Contacts between nucleotides upstream and downstream of the branch-site and the U2 SF3a/b proteins Prp9, Prp11, Hsh49, Cus1 and Hsh155 were detected, demonstrating that these interactions are evolutionarily conserved. The RES proteins Pml1 and Bud13 were shown to contact the intron downstream of the branch-site. A comparison of the Bact crosslinking pattern versus that of B* and C complexes revealed that U2 and RES protein interactions with the intron are dynamic. Upon step 1 catalysis, Cwc25 contacts with the branch-site region, and enhanced crosslinks of Prp8 and Prp45 with nucleotides surrounding the branch-site were observed. Cwc25's step 1 promoting activity was not dependent on its interaction with pre-mRNA, indicating it acts via protein-protein interactions. These studies provide important insights into the spliceosome's protein-pre-mRNA network and reveal novel RNP remodeling events during the catalytic activation of the spliceosome and step 1 of splicing.

Details

show
hide
Language(s): eng - English
 Dates: 2015-09-22
 Publication Status: Published online
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1371/journal.pgen.1005539
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: PLoS Genetics
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: 27 Volume / Issue: 11 (9) Sequence Number: e1005539 Start / End Page: - Identifier: -